Implantable cell chamber device and uses thereof

Abstract

To provide a suitable means for delivering a therapeutic biomolecule to a patient, wherein a stable and therapeutically effective blood level of the biomolecule occurs in a stable and convenient form over an extended period of time.SOLUTION: Provided is a device for treating a disease in a subject, wherein the subject may have Crohn's disease, ulcerative colitis, primary sclerosing cholangitis, eosinophilic esophagitis, autoimmune hepatitis, the device comprising a cell chamber comprising a population of cells that secrete a biomolecule for delivery to the subject.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Related Applications This application is a continuation of U.S. Provisional Application No. 62 / 991,422, filed March 18, 2020. The entire contents of the aforementioned priority application are incorporated herein by reference. can be.

[0002] Sequence Listing This application contains a Sequence Listing which has been submitted electronically in ASCII format, the entire contents of which are hereby incorporated by reference. The ASCII copy was created on March 11, 2021. ,T103022_1130WO_0465_1_SL.txt,66,553 The size in bytes.

[0003] The present invention relates to an implantable cell chamber device and a method for the delivery of therapeutic biomolecules. It relates to the relevant use for doing so on the subject. [Background technology]

[0004] With advances in biotechnology, many species of bacteria are being developed for pharmaceutical use using recombinant DNA technology. It has become possible to produce a variety of therapeutic biomolecules, such as proteins. Typically, each bolus dose is delivered by subcutaneous injection or intravenous infusion at regular intervals. The bioavailability of a molecule during administration is a function of its rates of absorption, distribution, and elimination. Bioavailability typically peaks shortly after administration and then declines gradually. The pharmacokinetic (PK) profile of each biomolecule generally varies between toxicity and minimal efficacy. The interval between doses is designed to maintain plasma drug concentrations within the therapeutic window. The need for frequent administration can be costly and inconvenient for patients. In addition, the PK profile and dosing frequency of each biomolecule will determine the drug concentration in each patient and the doses administered. Depending on the amount of biomolecule present in the dose and the time interval between doses, the concentration can range from very high to very low. The concentration is varied between low and high.

[0005] Because biomolecules can be larger and more complex than traditional inorganic drugs (i.e. The formulation of such biomolecules is particularly challenging. For example, in order for a protein to maintain biological activity, the formulation must , while preserving the conformational integrity of at least one core sequence of amino acids of the protein. Sometimes multiple functional groups of a protein must be protected from degradation. These compounds can suffer from a lack of stability during development, particularly in monoclonal antibodies and polyclonal antibodies. Reclonal antibodies can be relatively unstable (see, e.g., Wang, et al., JP Harm Sci.96:1-26(2007)). Chemical or physical properties of therapeutic proteins. Consequences of chemical instability include a reduction in the effective dose, e.g., due to irritation or immune reactivity. These include reduced safety of the therapy due to its short shelf life and more frequent manufacturing. Many formulation options are available, and one approach or system may be suitable for all purposes. It is not good for protein.

[0006] Therefore, there is a need to find suitable means for delivering therapeutic biomolecules to patients. This allows for stable blood levels of therapeutically effective biomolecules over a long period of time. It occurs in a consistent and convenient form. Summary of the Invention

[0007] Provided herein are devices capable of retaining cells while simultaneously providing a This device is a cell chamber device that allows the exchange of biomolecules between the cell chamber device and the outside. can be used, for example, to culture cells that perform functions including the secretion of therapeutic biomolecules When the cell chamber device is implanted in a subject, the device provides stable recordings over a long period of time. Additionally or alternatively, a chamber for biomolecule delivery can be provided to the subject. The cells within the device interact with the tissue surrounding or adjacent to the device and with the cells that grow within the device. The exchange of components between cells allows physiological functions such as removing toxins or performing metabolic operations. The chamber is capable of performing most or all of the biological functions in a mammalian host. It induces no fibrotic response, elicits little or no host immune response, and The cells are retained inside the chamber without leakage into the host tissue, and the host cells are allowed to enter the chamber. and / or therapeutic administration of biomolecules to a host, e.g., into the bloodstream. The scaffold comprises a layered structure composed of materials that allow for the delivery of a large amount of

[0008] Thus, in one aspect, provided herein is a multi-layered foot surrounding a cell chamber. a device including a cell chamber, the multi-layer scaffold including an outer layer and an inner layer in contact with the cell chamber; In some embodiments, the outer and inner layers each comprise a nanofibrous polymer. In some embodiments, the nanofibrous polymer can include, for example, nanofibrous polyester.

[0009] In some embodiments, the outer layer is made of nanofibrous polyethylene terephthalate, or In some embodiments, the outer layer comprises nanofibrous polybutylene terephthalate. It includes a mixture of ethylene terephthalate and polybutylene terephthalate. In an embodiment, the outer layer is made of electrospun polyethylene terephthalate, electrospun Polybutylene terephthalate, or electrospun polyethylene terephthalate and poly For example, nanofibrous polymethylene terephthalate. Other suitable polymers are described herein, including cellulose, or nanofibrous polyurethane. can be.

[0010] In some embodiments, the inner layer comprises nanofibrous polyurethane. The inner layer comprises electrospun polyurethane.

[0011] In some embodiments, the inner layer is made of nanofibrous polyethylene terephthalate, nanofibrous polyethylene terephthalate, Polybutylene terephthalate, or nanofibrous polyethylene terephthalate and polybutylene Includes mixtures with ethylene terephthalate.

[0012] In some embodiments, the inner layer is made of electrospun polyethylene terephthalate, Trospun polybutylene terephthalate, or electrospun polyethylene terephthalate and polybutylene terephthalate.

[0013] In some embodiments, the outer layer and / or the inner layer comprise one or more charged surface modifications.

[0014] In some embodiments, the outer layer and / or the inner layer have a net positive charge. In an embodiment, the scaffold is treated with ethylenediamine.

[0015] In some embodiments, the outer layer and / or the inner layer have a net negative charge. In an embodiment, the scaffold is treated with sodium hydroxide.

[0016] In some embodiments, the outer layer and / or the inner layer comprises an anti-inflammatory agent. In one embodiment, the anti-inflammatory agent is a calcineurin inhibitor. In some embodiments, the anti-inflammatory agent is a pyridone. In one embodiment, the anti-inflammatory In one embodiment, the anti-inflammatory agent is a phosphodiesterase inhibitor. In one embodiment, the anti-inflammatory agent is roflumilast.

[0017] In some embodiments, the outer layer and / or the inner layer comprise nanopores. In certain embodiments, the nanopores are sized to allow the passage of biomolecules. In some embodiments, the biomolecule is 250 kilodaltons (kDa) or less.

[0018] In some embodiments, the nanopore allows passage of an antibody, or antigen-binding portion thereof. In some embodiments, the nanopores are sized to prevent the passage of cells. In some embodiments, the nanopores allow cells on one side of the multilayer scaffold to penetrate the multilayer scaffold. The membrane is sized to prevent contact with cells on the other side of the membrane.

[0019] In some embodiments, the nanopore has a diameter of 1 μm or less. The nanopore has a diameter of 0.5 μm or less.

[0020] In some embodiments, the multi-layer scaffold comprises a nanoporous membrane positioned between an inner layer and an outer layer. In some embodiments, the nanoporous membrane is a solid, i.e., non-nanofiberous polymer. In other embodiments, the nanoporous membrane comprises an additional layer of nanofibrous polymer. In some embodiments, the nanoporous membrane comprises polybutylene terephthalate. In some embodiments, the nanoporous membrane is a solid, i.e., non-nanofiber polybutylene In certain embodiments, the nanoporous membrane comprises a layer of nanofibrous poly(ethylene terephthalate). In some embodiments, the nanoporous membrane comprises butylene terephthalate. In certain embodiments, the biomolecule comprises a membrane nanopore sized to allow It is less than 250 kDa.

[0021] In some embodiments, the membrane pores are configured to allow passage of antibodies, or antigen-binding portions thereof. In some embodiments, the membrane pores are sized to prevent the passage of cells. In some embodiments, the membrane pores allow cells on one side of the multi-layer scaffold to pass through to the other side of the multi-layer scaffold. It is sized to prevent contact with the cells.

[0022] In another aspect, provided herein is a device comprising a multi-layer scaffold surrounding a cell chamber. The multi-layer scaffold is a device comprising: (i) nanofibrous polyethylene terephthalate and polybutadiene; (ii) an outer layer comprising ethylene terephthalate; and (iii) a non-nano-based polymer positioned between the inner and outer layers. A fibrous membrane, the membrane being submicron in size to prevent the passage of cells across the membrane. and (iii) a non-nanofiberous membrane containing nanopores and / or nanofibers, and (iv) a nanofiberous polyurethane membrane. and an inner layer comprising:

[0023] In a further aspect, provided herein are multi-layered scaffolds surrounding a cell chamber. The device, comprising a multi-layer scaffold comprising: (i) nanofibrous polyethylene terephthalate and poly(ethylene terephthalate); (ii) an outer layer comprising butylene terephthalate; and (iii) a non-naphthalene rubber layer positioned between the inner and outer layers. a fibrous membrane, the membrane having nanopores of a size such as to prevent passage of cells across the membrane; and (iii) a non-nanofibrous membrane comprising nanofibrous polyethylene terephthalate and polybutadiene. and an inner layer comprising polyethylene terephthalate.

[0024] In some embodiments, the membrane nanopore has a diameter of 1 μm or less. In some embodiments, the membrane nanopores have a diameter of about 0.5 μm or less. In some embodiments, the membrane pores have a diameter of about 0.4 μm. do.

[0025] In some embodiments, the device includes an attachment that allows cells to be loaded into the cell chamber. A fill port is further provided.

[0026] In some embodiments, the device comprises a total thickness of 250 μm or less. The device comprises a total thickness of 150 μm or less.

[0027] In some embodiments, the cell chamber contains up to 1 x 10 9 A size large enough to accommodate 100 cells In some embodiments, the cell chamber contains up to 1 x 10 7 It can accommodate 100 cells. It's a large size.

[0028] In another aspect, provided herein is a device comprising a multi-layer scaffold surrounding a cell chamber. The multilayer scaffold is a device comprising nanofibrous polyethylene terephthalate and polybutylene an outer layer comprising terephthalate and an inner layer comprising nanofibrous polyurethane, The layer contains nanopores having diameters of 1 μm or less.

[0029] In some embodiments, the device is a scaffold having a thickness of 1 μm or more positioned between the outer and inner layers of the scaffold. The polymer film has the following pore size:

[0030] In some embodiments, the device comprises a polymer film having a pore size of 1 μm or less. One side of the film is coated with the first electrospun polymer, and the other side is coated with the second electrospun polymer. The other side of the film is coated with a second electrospun polymer. In this embodiment, the first electrospun polymer and the second electrospun polymer are the same. In another embodiment, the first electrospun polymer and the second electrospun polymer are The trospan polymer comprises a different polymer. The spun polymer and / or second electrospun polymer may be nPET, nPBT, or a combination thereof. In some embodiments, the first electrospun polymer In some embodiments, the first and / or second electrospun polymer comprises nPU. The first electrospun polymer included nPET-PBT, and the second electrospun polymer included nPET-PBT. The polymer includes nPU.

[0031] In some embodiments, the polymer film comprises poly(ethylene terephthalate). In embodiments, the polymer film includes pores having diameters of about 0.2 μm to 1 μm. In various embodiments, the polymer film has a thickness of about 0.2 μm, 0.3 μm, 0.4 μm, 0.5 μm, 1 μm, 2 μm, 3 μm, 4 μm, 5 μm, 6 μm, 7 μm, 8 μm, 9 μm, 10 μm, 11 μm, 12 μm, 13 μm, 14 μm, 15 μm, 16 μm, 0.5μm, 0.6μm, 0.7μm, 0.8μm, 0.9μm, or 1.0μm diameter In some embodiments, the polymer film comprises pores having a diameter of about 0.4 μm. The porous material includes pores having a thickness of 100 μm or less.

[0032] In some embodiments, the cell chamber comprises cells. In some embodiments, the cells are In some embodiments, the cells comprise retinal pigment epithelial cells. In some embodiments, the cells comprise ARPE-19 cells. In some embodiments, the cells comprise pancreatic islets.

[0033] In some embodiments, the device has a capacitance of about 1×10 5 cells ~ approx. 1 x 10 8 Contains 100 cells nothing.

[0034] In some embodiments, the device has a capacitance of about 1×10 6 cells ~ approx. 1 x 10 7 Contains 100 cells nothing.

[0035] In some embodiments, the cells secrete the recombinant peptide or protein. In this embodiment, the cells are provided with growth factors, hormones, cytokines, prostaglandins, enzymes, The protein secreted by the β-glucanase is selected from the group consisting of: a β-glucanase, ...

[0036] In some embodiments, the cells are administered as peptide therapeutics (e.g., gastrointestinal therapeutics, such as for the treatment of short bowel syndrome). The peptide therapeutic agent for use in the treatment of cancer is secreted.

[0037] In some embodiments, the cells secrete the enzyme. In one embodiment, the enzyme is In one embodiment, the enzyme is arylsulfatase A. In one embodiment, the enzyme is laronidase.

[0038] In some embodiments, the cells secrete antibodies, or antigen-binding portions thereof. In embodiments, the cells secrete the chimeric antibody, or an antigen-binding portion thereof. In some embodiments, the cells secrete the humanized antibody, or an antigen-binding portion thereof. In some embodiments, the cells secrete human antibodies, or antigen-binding portions thereof. In some embodiments, the cells secrete Fab, F(ab'), and monoclonal antibodies. 2. scFv, tandem scFv, diabody, minibody, and single domain antibody The antibody fragment is selected from the group consisting of:

[0039] In some embodiments, the cells express α4β7 integrin, integrin β7, TNFα, Specifically binds to an antigen selected from the group consisting of IL-12, IL-23, or CD20 In some embodiments, the cells secrete antibodies, or antigen-binding portions thereof, that inhibit the expression of VEGF. Mab, abrilumab, adalimumab, etrolizumab lizumab, certolizumab, golimumab, usute Antibodies selected from kinumab, infliximab, rituximab, and natalizumab are isolated. To secrete.

[0040] In some embodiments, the cells are infected with an antibody that specifically binds to α4β7 integrin, or In certain embodiments, the cells secrete vedolizumab, or an antigen-binding portion thereof. The antigen-binding portion is secreted.

[0041] In some embodiments, the cell chamber contains a population of cells having a three-dimensional structure. In some embodiments, the population of cells having a three-dimensional structure is a tissue explant. The tissue is liver tissue, kidney tissue, or pancreatic tissue. The population of cells is organized as a spheroid. The cells include hepatocytes, liver cells, or pancreatic cells. The cells are organized around sinusoids or ducts.

[0042] In another aspect, provided herein is a method for delivering a biomolecule to a subject, the method comprising: A method comprising administering to a subject a device as described in the present invention, The cell chamber contains cells that secrete biomolecules.

[0043] In another aspect, provided herein are recombinant peptides or proteins. A method of delivering to a subject, the method comprising administering to the subject a device of the present disclosure. .

[0044] In another aspect, provided herein are methods for delivering an antibody, or an antigen-binding portion thereof, to a subject. The method comprises administering to a subject a device of the present disclosure.

[0045] In some embodiments, the device is implanted under the skin (subcutaneous implantation), on the omentum, in the liver, in the brain, or It is administered to the subject by implantation at a selected site within the spinal canal.

[0046] In some embodiments, the device is implanted in the subject for at least 30 days. In some embodiments, the device is implanted in the subject for at least 90 days. The device is implanted in the subject for at least 120 days.

[0047] In some embodiments, the device is implanted in the subject for at least one year.

[0048] In some embodiments, the plasma concentration of the biomolecule, recombinant peptide, or recombinant protein is at least 5 μg / mL in the subject for a period of at least 60 days after implantation.

[0049] In some embodiments, the plasma concentration of the antibody, or antigen-binding portion thereof, is at least Both are at least 5 μg / mL in subjects for a 60-day period.

[0050] In another aspect, provided herein is a method for treating patients with Crohn's disease or ulcerative colitis. A method of treating an elephant comprising administering to the subject a device of the present disclosure.

[0051] In another aspect, provided herein are methods for treating a subject with primary sclerosing cholangitis. The method comprises administering to a subject a device of the present disclosure.

[0052] In another aspect, provided herein is a method for treating a subject with eosinophilic esophagitis. The method comprises administering a device of the present disclosure to a subject.

[0053] In another aspect, provided herein is a method for treating a subject with autoimmune hepatitis. The method comprises administering a device of the present disclosure to a subject.

[0054] In another aspect, provided herein is a method of treating a subject with short bowel syndrome. and a method comprising administering a device of the present disclosure to a subject.

[0055] In another aspect, provided herein is a method for treating patients with mucopolysaccharidosis type I (MPS I). A method of treating an elephant, comprising administering to the elephant a therapeutic agent ... In certain embodiments, the method includes administering to a subject a device that secretes a The device may be configured to detect an enzyme comprising the amino acid sequence of SEQ ID NO:50 or a low level of the amino acid sequence of SEQ ID NO:50. enzymes having at least 90%, 92%, 94%, 95%, 96%, 98%, or 99% Contains cells that secrete

[0056] In another aspect, provided herein is a method for treating a patient with mucopolysaccharidosis type II (MPS II). The present invention provides a method of treating a subject having a tumor comprising administering to the subject ... The method includes administering to a subject a device that secretes an enzyme, such as an enzyme. In embodiments, the device comprises an enzyme comprising the amino acid sequence of SEQ ID NO: 51, or At least 90%, 92%, 94%, 95%, 96%, 98%, or 99% of 1 The present invention includes cells that secrete an enzyme having the formula:

[0057] In another aspect, provided herein is a method for treating a patient with metachromatic leukodystrophy (MLD). The present invention provides a method of treating a subject with a cell-associated inflammatory cytokine, comprising administering to the subject ... The method includes administering to a subject a device that secretes an enzyme, such as taurateurase A. In certain embodiments, the device comprises an enzyme comprising the amino acid sequence of SEQ ID NO: 52, or the sequence At least 90%, 92%, 94%, 95%, 96%, 98% for number 52; or Contains cells that secrete enzymes that make up 99% of the

[0058] In another aspect, provided herein is a device comprising a monolayer scaffold surrounding a cell chamber. Such cell chamber devices are, in some embodiments, suitable for use in three-dimensional culture. It can be used to culture growing cells or as tissue explants. In embodiments, the cell chamber can contain cells having a three-dimensional structure. In embodiments, the monolayer scaffold may comprise a nanofibrous polymer as described herein. In some embodiments, the nanofibrous polymer is a polyester, e.g., a nanofibrous Polyethylene terephthalate or nanofibrous polybutylene terephthalate. In one embodiment, the single layer scaffold comprises nanofibrous polyethylene terephthalate and polybutylene terephthalate. In some embodiments, the single layer scaffold comprises a mixture of nanofibrous polyurethane and methyl phthalate. Includes

[0059] In some embodiments, the cell chamber contains cells having a three-dimensional structure. In some embodiments, the cells having a three-dimensional structure are tissue explants. In some embodiments, the tissue is liver. In another embodiment, the cell having a three-dimensional structure is a human tissue, a kidney tissue, or a pancreatic tissue. In some embodiments, the spheroids are organized as hepatocytes, liver In certain embodiments, the spheroids may comprise: In some embodiments, the cells are hepatocytes. In embodiments, the cells are pancreatic islet cells. [Brief explanation of the drawings]

[0060] [Figure 1A]1 shows scanning electron micrographs of native extracellular matrix, implantable woven polyester membrane, and electrospun polyester membrane. [Figure 1B] 1 shows confocal images of electrospun nanofibrous polyurethane (nPU) polymer (as part of an nPET-PBT / PU scaffold) 2 days after seeding the nPU layer with human ARPE-19 cells. [Figure 1C] 1 shows an exemplary cell chamber device. The cell chamber includes a bilayer scaffold surrounding the cell chamber. The bilayer scaffold includes an outer outward-facing layer (nPET-PBT) (top left) comprising electrospun nanofibrous polyethylene terephthalate (PET) and polybutylene terephthalate (PBT), and an inner inward-facing layer (bottom left) comprising electrospun nanofibrous polyurethane (nPU) polymer. In an exemplary embodiment, two scaffold sheets, or one folded sheet, can be fabricated to surround the cell chamber (top right). [Figure 1D] Figure 1 shows one embodiment of a multi-scaffold device. The device contains a first bilayer scaffold comprising an outer, outward-facing layer of electrospun nPET-PBT and an inner, inward-facing layer of electrospun nPU. In addition, the device contains a second scaffold comprising a porous layer of electrospun nanofibrous PBT. ARPE-19 cells seeded within the cell chamber can be visualized by hematoxylin and eosin (H&E) staining (bottom right). [Figure 1E] 1 shows one embodiment of a multilayer scaffold device comprising an outer, outward-facing layer of electrospun nPET-PBT, a central poly(ethylene terephthalate) film with a preselected pore size, and an inner, inward-facing layer of electrospun nPU. A diagram of the cell chamber device is shown in the left panel, and a schematic of the cell chamber device is shown in the right panel. [Figure 1F] 1C shows scanning electron micrographs of each layer in the cell chamber device shown in FIG. 1E. [Figure 1G]Figure 1 shows an image of a cell chamber containing an outer outward-facing layer of electrospun nPET-PBT, a central PET membrane with a 0.4 μm pore size, and an inner inward-facing layer of electrospun nPU. The cell chamber was seeded with ARPE-19 cells and visualized by hematoxylin and eosin (H&E) staining 44 days after cell seeding. [Figure 2] 1 shows a simulated model of the predicted plasma concentration of vedolizumab over time that can be achieved from a cell chamber device containing cells secreting 4.5 mg of vedolizumab / day or 9 mg of vedolizumab / day. The simulated model was based on the known pharmacokinetics of vedolizumab after subcutaneous injection. The therapeutically effective plasma concentration of vedolizumab is indicated by the bold line at 17 μg / mL. [Figure 3A] Figure 3A shows the results of an in vitro assay to assess the ability of vedolizumab-secreting ARPE-19 cells to grow on a scaffold consisting of nanofibrous polybutylene terephthalate (PBT) and polyethylene terephthalate (PET) on one side and a nanofibrous polyurethane (PU) polymer on the other side (nPET-PBT / PU scaffold). Figure 3A shows a fluorescence micrograph of vedolizumab / luciferase-ARPE-19 cells on a nPET-PBT / PU scaffold 24 hours after seeding on the inner PU side, taken from the PU side. [Figure 3B] Figure 3B shows the results of an in vitro assay to assess the ability of vedolizumab-secreting ARPE-19 cells to grow on scaffolds composed of nanofibrous polybutylene terephthalate (PBT) and polyethylene terephthalate (PET) on one side and nanofibrous polyurethane (PU) polymer on the other side (nPET-PBT / PU scaffolds). Figure 3B shows fluorescent micrographs of the nuclei (Hoescht 33342) or cytoplasm (CellTracker Orange) of vedolizumab / luciferase-ARPE-19 cells 8 days after seeding on normal tissue culture (TC) plates or nPET-PBT / PU scaffolds. [Figure 3C]Figure 3C shows the results of an in vitro assay to assess the ability of vedolizumab-secreting ARPE-19 cells to grow on scaffolds composed of nanofibrous polybutylene terephthalate (PBT) and polyethylene terephthalate (PET) on one side and nanofibrous polyurethane (PU) polymer on the other side (nPET-PBT / PU scaffolds). Figure 3C graphically depicts the amount of DNA isolated from vedolizumab / luciferase-ARPE-19 cells on normal tissue culture plates compared to nPBT-PET / PU scaffolds 40 days after seeding. [Figure 3D] Figure 3D shows the results of an in vitro assay to assess the ability of vedolizumab-secreting ARPE-19 cells to grow on scaffolds composed of nanofibrous polybutylene terephthalate (PBT) and polyethylene terephthalate (PET) on one side and nanofibrous polyurethane (PU) polymer on the other side (nPET-PBT / PU scaffolds). Figure 3D graphically depicts the amount of vedolizumab secreted by ARPE-19 cells on normal tissue culture plates (left bar) compared to nPBT-PET / PU scaffolds (right bar). [Figure 4A] Figure 4A graphically illustrates the results of an in vitro cell seeding assay of vedolizumab / luciferase-ARPE-19 cells seeded on nPET-PBT with charged surface modifications. Disks containing nPET-PBT / PU were treated with liquid ethylenediamine (EDA) to generate a positively charged surface or liquid sodium hydroxide (HYD) to generate a negatively charged surface. Figure 4A graphically illustrates the amount of vedolizumab secreted from ARPE-19 cells seeded on each of the indicated materials. [Figure 4B] Figure 4B graphically illustrates the results of an in vitro cell seeding assay of vedolizumab / luciferase-ARPE-19 cells seeded on nPET-PBT with charged surface modifications. Disks containing nPET-PBT / PU were treated with liquid ethylenediamine (EDA) to generate a positively charged surface or liquid sodium hydroxide (HYD) to generate a negatively charged surface. Figure 4B graphically illustrates the level of luminescence produced by vedolizumab / luciferase-ARPE-19 cells seeded on each of the indicated materials. [Figure 5A] Figure 5 shows the results of a study evaluating in vitro and in vivo cell distribution within a cell chamber containing an nPET-PBT / PU scaffold with or without an inner nPBT scaffold layer. Figure 5A shows luminescence images demonstrating the in vitro cell distribution of vedolizumab / luciferase-ARPE-19 cells growing on a cell chamber containing an nPBT-PET / PU scaffold with or without an inner nPBT scaffold. [Figure 5B] Figure 5B shows the results of a study evaluating in vitro and in vivo cell distribution within a cell chamber containing an nPET-PBT / PU scaffold with or without an inner nPBT scaffold layer. Figure 5B shows luminescence images of a mouse demonstrating the cell distribution of vedolizumab / luciferase-ARPE-19 cells growing within a cell chamber containing an nPBT-PET / PU scaffold with or without an inner nPBT scaffold after the indicated number of days after implantation. [Figure 5C] Figure 5C shows the results of a study evaluating in vitro and in vivo cell distribution within a cell chamber containing an nPET-PBT / PU scaffold with or without an inner nPBT scaffold layer. Figure 5C graphically depicts the luminescence intensity over time in mice implanted with vedolizumab / luciferase-ARPE-19 cells and a cell chamber containing an nPBT-PET / PU scaffold with or without an inner nPBT scaffold layer. [Figure 5D] Figure 5D shows the results of a study evaluating in vitro and in vivo cell distribution within cell chambers containing nPET-PBT / PU scaffolds with or without an inner nPBT scaffold layer. Figure 5D graphically depicts plasma concentrations (as detected by Western blot) over time in mice implanted with cell chambers containing vedolizumab / luciferase-ARPE-19 cells and PBT-PET / PU scaffolds with or without an inner nPBT scaffold layer. The bottom panel presents Western blot results detecting vedolizumab in plasma 68–78 days after implantation of the cell chambers. [Figure 5E]Figure 5E shows the results of a study evaluating in vitro and in vivo cell distribution within cell chambers containing nPET-PBT / PU scaffolds with or without an inner nPBT scaffold layer. Figure 5E shows luminescence images of a mouse 42 days after implantation before removal of the cell chamber (left column) and after removal of the cell chamber (right column). [Figure 5F] Results of a study evaluating in vitro and in vivo cell distribution within cell chambers containing nPET-PBT / PU scaffolds with or without an inner nPBT scaffold layer are shown. Figures 5F and 5G show images of nPBT-PET / PU cell chambers with an inner nPBT scaffold layer (Figure 5F; without cells (left column) or with cells (right column)) or without an inner nPBT scaffold layer (Figure 5G) after removal from nude mice 42 days after implantation, demonstrating that no significant fibrotic response was visually observed. [Figure 5G] Results of a study evaluating in vitro and in vivo cell distribution within cell chambers containing nPET-PBT / PU scaffolds with or without an inner nPBT scaffold layer are shown. Figures 5F and 5G show images of nPBT-PET / PU cell chambers with an inner nPBT scaffold layer (Figure 5F; without cells (left column) or with cells (right column)) or without an inner nPBT scaffold layer (Figure 5G) after removal from nude mice 42 days after implantation, demonstrating that no significant fibrotic response was visually observed. [Figure 5H] Figure 5H shows the results of a study evaluating in vitro and in vivo cell distribution within a cell chamber containing an nPET-PBT / PU scaffold with or without an inner nPBT scaffold layer. Figure 5H shows H&E stained images of the area surrounding a scaffold containing a porous non-nanofiber PET sheet coated with nanofibrous nPET-PBT and nPU (left panel) and the area surrounding a porous non-nanofiber PET sheet without the nanofibrous coating (right panel) 41 days after implantation in a mouse. [Figure 6]Figure 1 shows the results of an in vitro cell attachment assay, showing the percentage of adherent cells on discs made of nPET-PBT, nPET-PBT(ethylenediamine (EDA)), or nPET-PBT(sodium hydroxide (HYD)) compared to normal tissue culture (TC) plates as a function of time after cell seeding. 400,000 cells were seeded per disc. [Figure 7A] These figures show the results of an in vivo study in which vedolizumab / luciferase-ARPE-19 cell retention was assayed by quantifying luminescence intensity over time (days after implantation) in nude mice implanted with disks composed of nPET-PBT (Figures 7A, 7B, and 7E), positively charged nPET-PBT(EDA) (Figures 7A, 7C, and 7E), or negatively charged nPET-PBT(HYD) (Figures 7A, 7D, and 7E). Twenty-four or four hours after loading, the scaffolds were administered to mice by subcutaneous implantation (n = 4 mice). Cells administered to mice by subcutaneous injection were evaluated as a comparison. Figure 7A shows the results of the study over the course of 45 days. [Figure 7B] These figures show the results of an in vivo study in which vedolizumab / luciferase-ARPE-19 cell retention was assayed by quantifying luminescence intensity over time (days after implantation) in nude mice implanted with discs composed of nPET-PBT (Figures 7A, 7B, and 7E), positively charged nPET-PBT(EDA) (Figures 7A, 7C, and 7E), or negatively charged nPET-PBT(HYD) (Figures 7A, 7D, and 7E). Twenty-four or four hours after loading, the scaffolds were administered to mice by subcutaneous implantation (n = 4 mice). Cells administered to mice by subcutaneous injection were evaluated as a comparison. Figures 7B-7E show the results of the study over the course of 80 days. [Figure 7C]These figures show the results of an in vivo study in which vedolizumab / luciferase-ARPE-19 cell retention was assayed by quantifying luminescence intensity over time (days after implantation) in nude mice implanted with discs composed of nPET-PBT (Figures 7A, 7B, and 7E), positively charged nPET-PBT(EDA) (Figures 7A, 7C, and 7E), or negatively charged nPET-PBT(HYD) (Figures 7A, 7D, and 7E). Twenty-four or four hours after loading, the scaffolds were administered to mice by subcutaneous implantation (n = 4 mice). Cells administered to mice by subcutaneous injection were evaluated as a comparison. Figures 7B-7E show the results of the study over the course of 80 days. [Figure 7D] These figures show the results of an in vivo study in which vedolizumab / luciferase-ARPE-19 cell retention was assayed by quantifying luminescence intensity over time (days after implantation) in nude mice implanted with discs composed of nPET-PBT (Figures 7A, 7B, and 7E), positively charged nPET-PBT(EDA) (Figures 7A, 7C, and 7E), or negatively charged nPET-PBT(HYD) (Figures 7A, 7D, and 7E). Twenty-four or four hours after loading, the scaffolds were administered to mice by subcutaneous implantation (n = 4 mice). Cells administered to mice by subcutaneous injection were evaluated as a comparison. Figures 7B-7E show the results of the study over the course of 80 days. [Figure 7E] These figures show the results of an in vivo study in which vedolizumab / luciferase-ARPE-19 cell retention was assayed by quantifying luminescence intensity over time (days after implantation) in nude mice implanted with discs composed of nPET-PBT (Figures 7A, 7B, and 7E), positively charged nPET-PBT(EDA) (Figures 7A, 7C, and 7E), or negatively charged nPET-PBT(HYD) (Figures 7A, 7D, and 7E). Twenty-four or four hours after loading, the scaffolds were administered to mice by subcutaneous implantation (n = 4 mice). Cells administered to mice by subcutaneous injection were evaluated as a comparison. Figures 7B-7E show the results of the study over the course of 80 days. [Figure 8A]Figures 8A and 8B graphically show the results of an in vitro cell seeding assay in which vedolizumab / luciferase-ARPE-19 cells were seeded onto nPET-PBT loaded with tacrolimus (FK506). Discs containing nPET-PBT were treated with solutions containing 0%, 2%, or 4% tacrolimus. Figures 8A and 8B graphically show chromatograms from high-performance liquid chromatography (HPLC) analysis of tacrolimus alone (Figure 8A) or after extraction from tacrolimus-treated nPET-PBT (Figure 8B). [Figure 8B] Figures 8A and 8B graphically show the results of an in vitro cell seeding assay in which vedolizumab / luciferase-ARPE-19 cells were seeded onto nPET-PBT loaded with tacrolimus (FK506). Discs containing nPET-PBT were treated with solutions containing 0%, 2%, or 4% tacrolimus. Figures 8A and 8B graphically show chromatograms from high-performance liquid chromatography (HPLC) analysis of tacrolimus alone (Figure 8A) or after extraction from tacrolimus-treated nPET-PBT (Figure 8B). [Figure 8C] Figure 8C graphically illustrates the results of an in vitro cell seeding assay in which vedolizumab / luciferase-ARPE-19 cells were seeded onto nPET-PBT loaded with tacrolimus (FK506). Discs containing nPET-PBT were treated with a solution containing 0%, 2%, or 4% tacrolimus. Figure 8D graphically illustrates the results of a T cell activation assay in which T cell activation (measured by IL-1β production) was assessed after exposing human PBMCs to the medium surrounding tacrolimus-loaded nPET-PBT. [Figure 8D] Figure 8A shows the results of an in vitro cell seeding assay of vedolizumab / luciferase-ARPE-19 cells seeded on nPET-PBT loaded with tacrolimus (FK506). Discs containing nPET-PBT were treated with a solution containing 0%, 2%, or 4% tacrolimus. Figure 8D shows a fluorescence micrograph of vedolizumab / luciferase-ARPE-19 cells on nPET-PBT loaded with 0%, 2%, or 4% tacrolimus. [Figure 8E]Figure 8A and Figure 8B show graphs of the results of an in vitro cell seeding assay of vedolizumab / luciferase-ARPE-19 cells seeded on nPET-PBT loaded with tacrolimus (FK506). Disks containing nPET-PBT were treated with a solution containing 0%, 2%, or 4% tacrolimus. Figure 8E shows graphs of the amount of vedolizumab secreted from vedolizumab / luciferase-ARPE-19 cells seeded on nPET-PBT loaded with 0%, 2%, or 4% tacrolimus. [Figure 8F] Figure 8F graphically illustrates the results of an in vitro cell seeding assay of vedolizumab / luciferase-ARPE-19 cells seeded onto nPET-PBT loaded with tacrolimus (FK506). Discs containing nPET-PBT were treated with a solution containing 0%, 2%, or 4% tacrolimus. Figure 8F graphically illustrates the level of luminescence produced by vedolizumab / luciferase-ARPE-19 cells seeded onto nPET-PBT loaded with 0%, 2%, or 4% tacrolimus. [Figure 9A] The amount of antibody secretion from adalimumab / ARPE-19 cells (Figure 9A) or ustekinumab / ARPE-19 cells (Figure 9B) on normal tissue culture plates (left bar) compared to nPBT-PET / PU scaffolds (right bar) is shown graphically. [Figure 9B] The amount of antibody secretion from adalimumab / ARPE-19 cells (Figure 9A) or ustekinumab / ARPE-19 cells (Figure 9B) on normal tissue culture plates (left bar) compared to nPBT-PET / PU scaffolds (right bar) is shown graphically. [Figure 10A] 10A and 10B graphically depict the results of a functional assay to test the activity of adalimumab (FIG. 10A) or vedolizumab (FIG. 10B) secreted by ARPE-19 cells grown in serum-free medium. [Figure 10B] 10A and 10B graphically depict the results of a functional assay to test the activity of adalimumab (FIG. 10A) or vedolizumab (FIG. 10B) secreted by ARPE-19 cells grown in serum-free medium. [Figure 11A]Figure 1E shows the results of an in vitro study in which ARPE-19 cells stably expressing vedolizumab / luciferase were loaded into cell chambers at three different densities: 2.5 million, 5 million, and 10 million cells / chamber. The day after cell loading, the chambers were incubated in fresh Megavir SFM for 2 hours, followed by optical imaging using an IVIS Spectrum Imaging platform (PerkinElmer) by placing the chambers in medium containing 150 μg / ml D-luciferin. Figure 11A shows luminescence images of the nPET-PBT / PET / nPU cell chamber device (columns 1-3) or the Theracyte cell chamber device (Theracyte catalog number: PD20.0s; World Precision Instrument catalog number: 505396) (column 4) with the indicated cell numbers. Data were analyzed with Living Image software (PerkinElmer) by delimiting a fixed region of interest (ROI) around the device, quantifying total radiance in photons / second, and assessing the linearity of luminescence intensity versus cell number. [Figure 11B] Figure 11B shows the results of an in vitro study in which ARPE-19 cells stably expressing vedolizumab / luciferase were loaded into cell chambers at three different densities: 2.5 million, 5 million, and 10 million cells / chamber, as described in Figure 1E. The day after cell loading, the chambers were incubated in fresh Megavir SFM for 2 hours, followed by optical imaging using an IVIS Spectrum Imaging platform (PerkinElmer) by placing the chambers in medium containing 150 μg / ml D-luciferin. Figure 11B graphically depicts the luminescence intensity of the cells in the chambers shown in Figure 11A as a function of the number of cells in the chambers. [Figure 11C]Figure 11C shows the results of an in vitro study in which ARPE-19 cells stably expressing vedolizumab / luciferase were loaded into cell chambers at three different densities: 2.5 million, 5 million, and 10 million cells / chamber, as described in Figure 1E. The day after cell loading, the chambers were incubated in fresh Megavir SFM for 2 hours, followed by optical imaging using an IVIS Spectrum Imaging platform (PerkinElmer) by placing the chambers in medium containing 150 μg / ml D-luciferin. Figure 11C graphically depicts the concentration of vedolizumab in the medium as a function of cell number, as determined by a vedolizumab ELISA assay. [Figure 12A] The results of a study evaluating the in vivo expression of luciferase and / or vedolizumab by ARPE-19 cells in cell chambers as described in Figure 1E, including nPET-PBT / PU scaffolds with a central porous PET membrane (0.4 μm pores), were implanted into immunodeficient mice (nude mice; no T cells). Figure 12A shows luminescence images of nude mice implanted with nPET-PBT / PET / nPU cell chambers (top row) or Theracyte devices (bottom row). Data were analyzed with Living Image software (PerkinElmer) by demarcating a region of interest (ROI) around each device and quantifying total radiance in photons / second. [Figure 12B]Shown are the results of a study evaluating the in vivo expression of luciferase and / or vedolizumab by ARPE-19 cells in cell chambers as described in Figure 1E , containing nPET-PBT / PU scaffolds with a central porous PET membrane (0.4 μm pores), where the cell chambers were implanted in immunodeficient mice (nude mice; no T cells). FIG. 12B graphically depicts the average luminescence intensity over time in nude mice implanted with cell chambers as described in FIG. 1E containing vedolizumab / luciferase-ARPE-19 cells compared to the luminescence intensity in nude mice implanted with Theracyte devices containing vedolizumab / luciferase-ARPE-19 cells (Day 1-30: N=7; Day 31-60: N=5; Day 64: N=4 for nPET-PBT / PET / nPU, N=5 for Theracyte; Day 65-108: N=3 for nPET-PBT / PET / nPU, N=2 for Theracyte). [Figure 12C] Shown are the results of a study evaluating the in vivo expression of luciferase and / or vedolizumab by ARPE-19 cells in cell chambers as described in Figure 1E , containing nPET-PBT / PU scaffolds with a central porous PET membrane (0.4 μm pores), where the cell chambers were implanted in immunodeficient mice (nude mice; no T cells). Figure 12C graphically depicts the mean vedolizumab plasma concentration (as detected by ELISA) over time in nude mice implanted with cell chambers as described in Figure 1E containing vedolizumab / luciferase-ARPE-19 cells, compared to the vedolizumab plasma concentration over time in nude mice implanted with Theracyte cell chambers containing vedolizumab / luciferase-ARPE-19 cells (Days 1-30: N=7; Days 31-60: N=5; Day 64: N=4 for nPET-PBT / PET / nPU, N=5 for Theracyte; Days 65-108: N=3 for nPET-PBT / PET / nPU, N=2 for Theracyte). Horizontal lines indicate target therapeutic plasma concentrations. [Figure 12D]Figure 12D shows the results of a study evaluating the in vivo expression of luciferase and / or vedolizumab by ARPE-19 cells in cell chambers as described in Figure 1E containing nPET-PBT / PU scaffolds with a central porous PET membrane (0.4 μm pores), where the cell chambers were implanted in immunodeficient mice (nude mice; no T cells). Figure 12D graphically depicts the mean vedolizumab plasma concentration (detected by ELISA) over time in nude mice implanted with cell chambers as described in Figure 1E containing vedolizumab-ARPE-19 cells. The horizontal line indicates the target therapeutic plasma concentration. [Figure 12E] Figure 12E shows the results of a study evaluating the in vivo expression of luciferase and / or vedolizumab by ARPE-19 cells in a cell chamber as described in Figure 1E, which contained an nPET-PBT / PU scaffold with a central porous PET membrane (0.4 μm pores), implanted in an immunodeficient mouse (nude mouse; no T cells). Figure 12E contains H&E stained images (left panel) and Masson's trichrome stained images (right panel) of the area surrounding a cell chamber as shown in Figure 1E, which contained a non-nanofibrillar PET sheet (membrane) with 0.4 μm pores, coated on one side with electrospun nPET-PBT and on the other side with electrospun nPU, and seeded with ARPE-19 cells, 30 days (top panel) or 64 days (bottom panel) after implantation in the mouse. [Figure 12F] Figure 12F shows the results of a study evaluating the in vivo expression of luciferase and / or vedolizumab by ARPE-19 cells in cell chambers as described in Figure 1E, including nPET-PBT / PU scaffolds with a central porous PET membrane (0.4 μm pores), where the cell chambers were implanted in immunodeficient mice (nude mice; no T cells). Figure 12F contains an H&E stained image of the area surrounding an nPET-PBT / PET / nPU cell chamber (as shown in Figure 1E) without cells seeded therein, 60 days after implantation in mice. [Figure 12G]The results of a study evaluating the in vivo expression of luciferase and / or vedolizumab by ARPE-19 cells in cell chambers as described in Figure 1E, including nPET-PBT / PU scaffolds with a central porous PET membrane (0.4 μm pores), were implanted in immunodeficient mice (nude mice; no T cells). Figure 12G shows images of nPBT-PET / PET / nPU cell chambers (as shown in Figure 1E) or Theracyte cell chambers seeded with ARPE-19 cells expressing vedolizumab and luciferase after removal from nude mice 30 days (top panel) or 64 days (bottom panel) after implantation. [Figure 12H] Figure 12H shows the results of a study evaluating the in vivo expression of luciferase and / or vedolizumab by ARPE-19 cells in cell chambers as described in Figure 1E, containing nPET-PBT / PU scaffolds with a central porous PET membrane (0.4 μm pores), where the cell chambers were implanted in immunodeficient mice (nude mice; no T cells). Figure 12H shows luminescence images of a nude mouse before and after removal of a PBT-PET / PET / nPU cell chamber (as shown in Figure 1E) containing vedolizumab / luciferase-ARPE-19 cells, compared with luminescence images of a nude mouse before and after removal of a Theracyte cell chamber containing vedolizumab / luciferase-ARPE-19 cells. [Figure 13A] Graphical representations show the mean plasma concentrations over time of adalimumab (FIG. 13A; Days 1-60: N=5; Days 61-81: N=4; Days 82-92: N=3; Days 93-108: N=2) or ustekinumab (FIG. 13B; N=5) in immunodeficient mice implanted with cell chambers as described in FIG. 1E containing adalimumab / ARPE-19 cells (FIG. 13A) or ustekinumab / ARPE-19 cells (FIG. 13B). [Figure 13B]Graphical representations show the mean plasma concentrations over time of adalimumab (FIG. 13A; Days 1-60: N=5; Days 61-81: N=4; Days 82-92: N=3; Days 93-108: N=2) or ustekinumab (FIG. 13B; N=5) in immunodeficient mice implanted with cell chambers as described in FIG. 1E containing adalimumab / ARPE-19 cells (FIG. 13A) or ustekinumab / ARPE-19 cells (FIG. 13B). [Figure 14] 1 shows a workflow for an experiment testing the ability of intestinal-derived tissue explants to grow in a monolayer cell chamber device containing nanofibrous electrospun polymer. [Figure 15] Representative images of cell chamber devices loaded with intestinal tissue explants are presented before implantation in a rat model (top panel), 7 days after implantation (bottom left panel), and 28 days after implantation (bottom right panel). [Figure 16A] Representative images of cell chamber devices containing nanofibrous nPET-PBT loaded with roflumilast (FIG. 16A, top panel), pirfenidone (FIG. 16A, bottom panel), or tacrolimus (FIG. 16B) are presented. The devices were loaded with intestinal tissue explants prior to implantation in a rat model and imaged at 7 and 28 days after implantation. [Figure 16B] Representative images of cell chamber devices containing nanofibrous nPET-PBT loaded with roflumilast (FIG. 16A, top panel), pirfenidone (FIG. 16A, bottom panel), or tacrolimus (FIG. 16B) are presented. The devices were loaded with intestinal tissue explants prior to implantation in a rat model and imaged at 7 and 28 days after implantation. DETAILED DESCRIPTION OF THE INVENTION

[0061] I. Definition As used herein, the term "nanofiber" refers to fibers having a diameter of less than 3.0 micrometers. In an exemplary embodiment, nanofibers refer to fibers having a diameter of 10 nanometers to 3 The diameter of the nanofibers varies depending on the type of polymer used and It depends on the method of production. Methods for producing nanofibers include electrospinning, These include bonding, self-assembly, template synthesis, and thermally induced phase separation.

[0062] As used herein, the term "electrospinning" refers to the process of forming a crosslinked mesh. A process that uses high voltage in combination with a distance from the material source to the base to produce a For example, an electric field is used to drive a solution containing a polymer from the tip of a capillary. A high voltage DC current can be applied to the solution, which This allows the jet of solution to be drawn towards a grounded collector screen. Upon exiting the capillary orifice, the charged solution jet is vaporized and forms a fiber. A solution can be formed which is collected on a collector (e.g., a rotating collector). The size and shape of the fibers thus obtained can be determined by the viscosity of the solution, the molecular weight, , polymer properties, and other parameters related to the electrospinning device. The electrospinning process for forming polymer nanofibers depends on the following factors: It has been demonstrated using a variety of polymers (see, for example, Huang, et al., Composites Science and Technology (2003) 63:2223-2253).

[0063] As used herein, "electrospun polymer" refers to a polymer that is produced by electrospinning. Electrospun refers to polymer fibers, such as polymer nanofibers, produced by electrospun polymer nanofibers. The polymers are polyethylene terephthalate (PET), polybutylene terephthalate (P The materials may include, but are not limited to, polyethylene (PET), polyethylene terephthalate (PE), or polyurethane (PU). The polymers described above can be electrochemically synthesized to produce polymer nanofibers, as described herein. It can be cutrospun.

[0064] As used herein, the term "biomolecule" includes polypeptides, polysaccharides, and Any organic molecule that can be produced by a living organism, including polynucleotides, as well as lipids (e.g., phospholipids, glycolipids, and sterols), chemical messengers (e.g., hormones and neurotransmitters), vitamins, sugars (e.g., carbohydrates, disaccharides, oligosaccharides, polysaccharides), amino acids, peptides, oligopeptides, polypeptides, proteins, nucleotides, deoxyribonucleic acids It refers to organic molecules such as DNA (dinucleotide polynucleotides) or RNA (ribonucleic acid). In some embodiments, the biomolecule is recombinantly produced. by other means, such as extracellular vesicles, exosomes, or secretory organelles such as mitochondria. The secreted biomolecule may be secreted from the cell. For example, the secreted biomolecule may be an exosome, a lipid polymer, or This may include those assembled, packaged, and secreted as viral particles. In this embodiment, the biomolecule, when provided to a subject in an effective amount or dose, is effective to treat a disease, disorder, infection, or other condition. , or a subject in need of treatment, prevention, and / or amelioration of symptoms of a disease, e.g. In certain embodiments, the antibody may be a "therapeutic biomolecule" capable of doing so in a human patient. The cells grown in the cell chamber devices described herein are capable of absorbing the energy from the cells when the device is administered. In some embodiments, the therapeutic biomolecule can be secreted in an effective amount in the subject. In some embodiments, the therapeutic biomolecule is an antibody. In other embodiments, the therapeutic biomolecule is a hormone. In another embodiment, the therapeutic biomolecule is an enzyme. The molecule is a peptide or a protein.

[0065] The term "antibody" as used herein refers to an antibody that binds to other molecules via disulfide bonds. The immune system consists of four polypeptide chains, two heavy (H) chains, and two light (L) chains, connected together. Each heavy chain comprises a heavy chain variable region (abbreviated herein as HCVR or VH). The heavy chain constant region consists of three domains, CH Each light chain consists of a light chain variable region (herein referred to as LCVR or CH3). The light chain constant region consists of one domain, C The VH and VL regions are composed of more conserved regions called framework regions (FR). Interspersed with these regions are regions of hypervariability called complementarity-determining regions (CDRs), which further Each VH and VL consists of three CDRs and four FRs. From the amino terminus to the carboxy terminus, they are arranged in the following order: FR1, CDR1, FR2, CDR3, CDR4, CDR5, CDR6, CDR7, CDR8, CDR9, CDR10, CDR11, CDR12, CDR13, CDR14, CDR15, CDR16, CDR17, CDR18, CDR19, CDR11, CDR12, CDR13, DR2, FR3, CDR3, FR4. In some embodiments, the antibody is a fragment crystallizable (F Antibodies suitable for use in the embodiments described herein include IgG ( For example, IgG1, IgG2, IgG3, IgG4), IgM, IgA1, IgA2, I The antibody may include an antibody having an isotype selected from IgE, IgD, or IgE. In embodiments, the antibody is an IgG1 antibody, an IgG2 antibody, or an IgG3 antibody. In embodiments, the antibody may comprise a kappa or lambda light chain. The antibody may have an IgG1 isotype and a kappa light chain.

[0066] The cell surface molecule, "α4β7 integrin" or "α4β7" (interchangeably used throughout) (used for this purpose) is a heterozygote consisting of the α4 chain (CD49D, ITGA4) and the β7 chain (ITGB7). Human α4-integrin and β7-integrin genes, GenBan k(National Center for Biotechnology Info rmation, Bethesda, Md.) The reference sequence accession numbers are NM_00 and NM_10, respectively. 0885 and NM_000889) are B and T lymphocytes, especially memory CD4+ lymphocytes. Typical of many integrins, α4β7 is expressed by endothelial cells in either a resting or activated state. The ligand for α4β7 is vascular cell adhesion molecule (VCAM). ), fibronectin, and mucosal addressins (MAdCAMs (e.g., MAdCAM-1 Antibodies that bind to α4β7 integrin are referred to herein as "anti-α4β These antibodies are called "antibodies No. 7."

[0067] As used herein, an antibody having "binding specificity for the α4β7 complex" or or an antigen-binding fragment thereof binds to α4β7 but not to α4β1 or α E Does not bind to B7 Vedolizumab is an example of an antibody with binding specificity for the α4β7 complex.

[0068] The "CDRs" or "complementarity determining regions" are referred to as "framework regions" (FRs). , regions of hypervariability interspersed within more conserved regions.

[0069] As used herein, the term "antibody fragment" or "antigen-binding fragment" of an antibody , Fab, Fab', F(ab')2, and Fv fragments, single chain antibodies, functional heavy chain antibodies (na an antibody), and at least one desired antibody that competes with the intact antibody for specific binding. Any portion of an antibody that has specificity for an epitope (e.g., a portion that specifically binds to the epitope) refers to an isolated portion of a complementarity-determining region (an isolated portion of a complementarity-determining region having sufficient framework sequence to match the complementarity-determining region of a given gene) Antigen-binding fragments can be produced by recombinant techniques or by enzymatic or chemical cleavage of the antibody. Exemplary antibody fragments include Fab, F(ab')2, scFv, and tandem These include dem-scFv, diabodies, minibodies, and single domain antibodies. Not limited to.

[0070] "Humanized" forms of non-human (e.g., rodent) antibodies contain minimal sequence derived from the non-human antibody. For the most part, humanized antibodies are chimeric antibodies containing the Residues from the hypervariable regions are selected from mouse, rat, and rabbit antibodies with the desired specificity, affinity, and potency. by residues from the hypervariable regions of a non-human species (donor antibody), such as a non-human primate, or a non-human primate. In some cases, the recipient antibody is replaced by a human immunoglobulin (recipient antibody). In this example, framework region (FR) residues of the human antibody are replaced by corresponding non-human residues. Furthermore, humanized antibodies contain residues that are not found in either the recipient antibody or the donor antibody. These modifications may be made to further refine antibody performance. Generally, human A humanized antibody is one in which all or substantially all of the hypervariable CDR loops are hypervariable CDR loops of a non-human antibody. corresponding to a sequence of the FRs, wherein all or substantially all of the FRs are those of a human immunoglobulin sequence; It will contain substantially all of at least one, and typically two, variable domains. The antibody optionally comprises at least an antibody constant region (Fc), typically the constant region of a human antibody. For further details, see, for example, Jones et al. ,Nature321:522-525(1986), Riechmann et al. .,Nature332:323-329(1988), and Presta,Curr. Op.Struct.Biol.2:593-596(1992).

[0071] As used herein, the term "pore(s)" refers to the flow of material from one side to the other. Pores refer to openings in a material, such as a scaffolding material, that allow the passage of biomolecules. It may be of any shape or size suitable for this purpose. In the case of electrospun nanofibrous polymers, the spaces between the fibers function as pores. In one embodiment, the pores allow the passage of biomolecules but not cells. The conductor may be sized to block the

[0072] As used herein, the term "recombinant protein" refers to a protein produced in a host cell, e.g. Gene(s) carried by recombinant expression vector(s) introduced into mammalian host cells ) refers to a protein produced as a result of transcription and translation. The recombinant protein is a recombinant antibody, or an antigen-binding portion thereof.

[0073] The term "recombinant host cell" (used interchangeably herein with the term "host cell") (e.g., a recombinant expression vector encoding a secreted protein) Such terms include not only specific target cells but also cells into which a vector has been introduced. It should be understood that the term "mutation" or "environment" is intended to refer to the progeny of such cells. Certain modifications can occur in subsequent generations due to the influence of either Although a cell may not, in fact, be identical to the parent cell, it is still considered a "cell" as used herein. Furthermore, unless otherwise specified, the term "cell" shall be used to refer to a cell that is a host cell. When terms such as host cell or mammalian cell or mammalian host cell are used, the term refers to a cell. It should be understood that the term is intended to include populations of cells.

[0074] The term "about" means that the value that follows is not an exact value, but rather a range of ±5% of the value. If the value is a relative value given as a percentage, then "approximately" is used. The term " " also indicates that the value that follows is not an exact value, but the center point of a range that is ±5% of the value. whereby the upper limit of the range cannot exceed the value of 100%.

[0075] II. Implantable Cell Chamber Device Provided herein are methods for preserving cells, e.g., cells that secrete therapeutic biomolecules. The cell chamber device of the present disclosure is a cell chamber device capable of The scaffold includes a nanofibrous polymer that defines an envelope of the protein. It allows the passage of biomolecules, such as proteins, into and out of the cell chamber, but does not allow the passage of cells. Living cells capable of producing biomolecules can be loaded into the cell chamber, Here, cells can grow and adhere to the inner surface of the scaffold. Thus, secreted biomolecules can diffuse out of the cell chamber through the pores in the scaffold. The scaffold prevents the cells contained therein from migrating out of the cell chamber, allowing the host cells to When the cell chamber is implanted into a host subject, the cells The cells within the chamber device allow for the stable delivery of various biomolecular therapeutics in stable forms. For example, it can be provided for the treatment of a host subject in need thereof. The continuous production of a desired biomolecule by the cells can be achieved by conventional means by preparing an isolated culture medium containing the biomolecule. The cells provided herein can be used to treat a variety of conditions, including the development of a tumour, a cytoplasm, a cytoplasmic or cytoplasmic nucleoside analogue, or a cytoplasmic or cytoplasmic nucleoside analogue. The chamber device preferably causes little or no fibrosis in a mammalian host. Inducing no response and including materials that induce little or no host immune response.

[0076] The cell chamber devices provided herein include a nanofibrous porous membrane surrounding the cell chamber. In some embodiments, the scaffold of the cell chamber device comprises a multi-layer structure. For example, the scaffold may include two layers of nanofibrous polymer having an outer layer and an inner layer. For example, the outer layer of the nanofibrous polymer may be in contact with the external environment surrounding the device. Alternatively, the scaffold may have an inner layer of nanofibrous polymer in contact with the cell chamber. By including one or more additional layers between the inner layer, a three-layer or multi-layer structure can be obtained. For example, the scaffold may optionally include a membrane, e.g., a porous membrane, positioned between an outer layer and an inner layer. The membrane may alternatively be referred to herein as a porous membrane, semi-porous membrane, or non-porous membrane. In some embodiments, one or more additional layers of the scaffold are nanofibrous. In some embodiments, the multilayer scaffold comprises 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more layers. The membrane layers (e.g., the central membrane layer) may comprise the same nanofibrous polymer or, alternatively, may comprise different For example, the outer layer of the scaffold may comprise different nanofibrous polymers with properties that are specific to the host. Suitable for implantation within tissue (e.g., most or all tissues within the implant recipient) does not generate any fibrotic or immune response and allows vascularization of the outer scaffold) The inner layer that directly contacts the cells in the cell chamber can contain the cell paws. The material may include materials that can function as a field and optionally a barrier. In this embodiment, the inner layer may have smaller pores than the outer layer.

[0077] The inner and / or outer scaffold layers of the cell chamber device may be polyethylene terephthalate. (PET, also known as Dacron), Polybutylene Terephthalate (PBT ), or polyurethane (PU), or various nanofibrous polymers. Any biocompatible polymer suitable for use in electrospinning may be used as described herein. In some embodiments, the cell chamber device may be made of a biocompatible polymer. The cellulose membrane is a nanofibrous polyester. Other polymers suitable for use in the scaffold layer include polylactic acid (PLA), polyglycolic acid (PGA), polylactic-co-glycolic acid (PLGA), polycaprolactone (PCL) , Polypropylene (PP), Polytetrafluoroethylene (PTFE), Polytrimethyl Polyethylene terephthalate, polyvinyl alcohol (PVA), polyethylene oxide (PEO ), Polytrimethylene Polyterephthalate (PTT), Polyethylene Acetate (PEV A), poly-D-lactide (PLDA), polylactic acid (PLLA), or polyethylene glycol Other suitable polymers include, but are not limited to, polyethylene glycol (PEG). Collagen, gelatin, alginate, fibrinogen, silk, elastin, cellulose The inner and / or outer layers may be made of, but are not limited to, cellulose, chitin, and chitosan. , each may comprise a single type of nanofibrous polymer. In such a case, the inner layer and / or Alternatively, the outer layers may each comprise a different type of nanofibrous polymer, or may comprise the same type of nanofibrous polymer. Alternatively, the inner and / or outer layers may each comprise a nanofibrous polymer of the type It may contain more than one type of nanofibrous polymer.

[0078] For example, in some embodiments, the outer layer may be made of nanofibrous polyethylene terephthalate (nP The polymers may include nanofibrous polybutylene terephthalate (nPBT) or nanofibrous polybutylene terephthalate (nPBT). In some embodiments, the outer layer is made of nanofibrous polyethylene terephthalate and polybutylene In some embodiments, the polymer may include both PET and PBT. The outer layer can include nanofibrous polyurethane (nPU).

[0079] In some embodiments, the inner layer is made of nanofibrous polyethylene terephthalate (nPET) or or nanofibrous polybutylene terephthalate (nPBT). In an embodiment, the inner layer is made of nanofibrous polyethylene terephthalate and polybutylene terephthalate. In some embodiments, the inner layer may comprise both a PET-PBT and a PET-PBT. In some embodiments, the outer layer may comprise a nanofibrous polyurethane (nPU). The inner layer may comprise nPET-PBT and the inner layer may comprise nPU. In some embodiments, the outer layer may comprise nPU. In some embodiments, the inner layer may comprise nPET, n It may contain PBT, or nPET-PBT.

[0080] The scaffold may additionally or alternatively comprise other synthetic or biological materials. Examples of materials include polytetrafluoroethylene (ePTFE) or poly(glycol). Biomaterials include biological membranes, bovine tissue, and collagen. Scaffolds (e.g., gels, threads, foams, sheets, mats, or tubes), gelatin, glutathione, cellulose (e.g., methylcellulose), elastin, glycosaminoglycans Examples of biomolecules include cellulose, peptidoglycan, chitin, or fibrin (e.g., gel). The material may optionally contain a cross-linking agent, such as an aldehyde (e.g., glutaraldehyde or formaldehyde), carbodiimides (e.g., 1-ethyl-3-(dimethylaminopropyl) carbodiimide), acrylamide (e.g., N,N'-methylenebisacrylamide), diimidates (e.g., dimethylsuberimidate) or diimidates (e.g., dimethylsuberimidate). In some embodiments, the biological material comprises a glutaraldehyde cross-linked biological membrane. include.

[0081] Synthesizing tissue scaffolds that can be used to fabricate the cell chamber devices described herein. There are several different methods for this. These include, for example, electrospinning, nanofibers, Examples of methods for fabricating scaffold layers include self-assembly, textile techniques, solvent casting, and particulate leaching. An exemplary method is electrospinning, which uses an electric field to spin polymers containing This is the process by which a solution is drawn from the tip of a capillary into a collector. A jet of solution is applied to the solution, which is directed towards a grounded collector screen. Once ejected from the capillary orifice, a charged liquid jet The is evaporated to form fibers, which are collected on a collector. using cross-linked meshes of polymer fibers with diameters ranging from nanometers to micrometers. The size and shape of the fibers obtained by electrospinning can be The morphology is related to the viscosity of the solution, the molecular weight, the nature of the polymer, and the electrospinning equipment. The electrochemical reaction for forming polymer nanofibers depends on various factors, including the amount of electrons and other parameters. The trospinning process has been demonstrated using a variety of polymers (e.g., all of which Huang, et al. Composite s Science and Technology(2003)63:2223-22 53).

[0082] In certain embodiments, the inner and / or outer layers of the scaffold are electrospun. The electrospun polymer is then produced, thereby forming a scaffold comprising the electrospun polymer. Rospan polymers are biocompatible and form structures similar to the body's natural extracellular matrix scaffolding. It has a random web-like structure, which promotes tissue integration and prevents rejection. Furthermore, electrospun polymers improve cell growth, promote cell differentiation, and facilitate cell attachment. In addition, the size of the electrospun fibers can be adjusted to accommodate other embedded Fibers of possible materials (e.g., woven polyester films, see Figures 1A-1C) It can be made 10 to 14 times smaller than the fiber size.

[0083] In an exemplary embodiment, the scaffold (e.g., inner and / or outer layers) is electrospun. Polyethylene terephthalate, electrospun polybutylene terephthalate, or The polymer may include an electrospun polymer selected from an electrospun polyurethane. In some embodiments, the outer layer comprises electrospun polyethylene terephthalate. In some embodiments, the outer layer comprises electrospun polybutylene terephthalate. In some embodiments, the outer layer is made of electrospun polyethylene terephthalate and It contains Trospan polybutylene terephthalate (see Figure 1C, bottom image). In one embodiment, the inner layer comprises electrospun polyurethane (FIG. 1C, top image). In an exemplary embodiment, the outer layer is made of electrospun polyethylene terephthalate. the inner layer comprises electrospun polybutylene terephthalate and electrospun polybutylene terephthalate; Includes spun polyurethane.

[0084] Electrospun polymers are known in the art or are further described herein. In one embodiment, the electrospanning procedure described herein can be used to generate Spun polymers are first prepared by mixing the polymer with hexafluoroisopropanol (HFIP). The polymer solution may be prepared by preparing a polymer solution, such as a polymer solution containing a polymer Ethylene terephthalate (PET) and polybutylene terephthalate (PBT) solutions To form the PET and PBT chips or pellets, place them in a solution of HFIP, Keep the chips in a cool, dry place (e.g., on a rotator) until the chips dissolve (e.g., incubation for approximately 5–7 days). Similarly, a solution of polyurethane (PU) can be formed. To do this, PU chips or pellets are placed in a solution of HFIP until the chips dissolve ( Incubate (e.g., on a rotator) for approximately 5-7 days. It can be done.

[0085] The polymer solution is then loaded onto the electrospinning unit and electrospun. For example, a polyester solution can be loaded into a syringe and This can then be connected to the electrospinning unit. The electrospinning unit was operated according to standard operating procedures to produce electrospun polyimide. In some embodiments, a polymer (e.g., PET, PBT, or PU) can be produced. In this case, the electrospinning unit should be approximately 10-25 cm (e.g., approximately 10 cm, approximately 1 1cm, approx. 12cm, approx. 13cm, approx. 14cm, approx. 15cm, approx. 16cm, approx. 17cm, Approximately 18cm, approximately 19cm, or approximately 20cm, approximately 21cm, approximately 22cm, approximately 23cm, approximately 24 cm, or approximately 25 cm) electrospinning distance, approximately 15-30 kV (e.g. , about 15 kV, about 18 kV, about 20 kV, about 21 kV, about 22 kV, or about 23 kV, an electrospinning voltage of about 25 kV, about 28 kV, or about 30 kV, or about 1 5-150 RPM (e.g., about 15-25 RPM, about 25-50 RPM, about 50-75 RPM) PM, approximately 75-100 RPM, approximately 100-125 RPM, or approximately 125-150 RPM In some embodiments, the second electrospun is set to have a rotation speed of 1 / 2 . The polymer (e.g., PU layer) is a first electrospun polymer (e.g., nPET- PBT layer) and consists of two or more layers of different electrospun polymers In certain embodiments, the electrospun material can be prepared by In another embodiment, the electrospun material contains nPU layered on nPET-PBT. Using such a bilayer scaffold, The cell chamber devices described herein can be fabricated.

[0086] One or more additional polymers are optionally layered on or between the aforementioned layers. and used to fabricate some embodiments of the cell chamber devices described herein. In some embodiments, a multi-layered scaffold can be formed that can: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more layers of nanofibrous polymer, e.g. The layers may each comprise a different polymer. or multiple layers can comprise the same polymer.

[0087] The cell chamber can be fabricated from multi-layer (e.g., bi-layer) electrospun polymer materials. The material can be measured and cut to any desired size. Once sized, the edges of the electrospun polymer material can be partially or completely sealed. The polymeric material can be formed into a chamber by methods known in the art. Thus, the edges of the chamber can be sealed. In one embodiment, the edges of the chamber are In some embodiments, at least one portion of the chamber is sealed using sonic welding. are left unsealed, for example to allow subsequent loading of cells. In this embodiment, the chamber is completely sealed, but can later be opened, e.g., by cutting. a cell loading port that can be opened by piercing or by needle piercing and subsequently sealed In some embodiments, the scaffold material is folded before sealing the edges. In some embodiments, a cell loading port (e.g., attached to a cell injection device) The port (with a Luer lock mechanism that allows cell loading) is designed to allow cell loading. After loading, the port is optionally removed and the chamber is ultrasonically The chamber is sealed by wave or heat welding, or by sealing the chamber opening with an adhesive. It can be sealed.

[0088] In some cases, the inner layer of the scaffold and / or the cell chamber device herein Alternatively, the nanofibrous polymer comprising the outer layer is further modified with one or more charged surface modifications. For example, in some embodiments, nanofibrous structures in the inner and / or outer layers of the scaffold can be The polymer is contacted with liquid ethylenediamine. For example, when reacted with PET, the ethylenediamine Treatment with benzodiamine results in a positively charged surface. The nanofibrous polymer of the inner and / or outer layer is contacted with liquid sodium hydroxide. When reacted with PET, for example, by treatment with sodium hydroxide, the negatively charged surface Altering the surface charge of a material can be used to bind to cell surface proteins. Charged surface materials can also modulate the attraction or adhesion of cell types that have a specific charge. It attracts specific proteins, such as those in the blood and the surrounding environment, which promote cell attachment. Ionic surfaces can also improve surface wettability and cell contact. Charged surface modifications can be cationic functional groups, anionic functional groups, or cationic functional groups. For example, the charged surface modification may include both carboxylic acid and anionic functional groups. In some embodiments, the charged surface modification can include a scaffold having a surface functional group or a surface functional group. by treating the polymer with alkaline hydrolysis (e.g., sodium hydroxide treatment) In some embodiments, the charged surface modification can be generated by - Ethylenediamine (EDA), 2-methylpentamethylenediamine, 1,2-diamin By treating with diamines such as 1,6-hexanediamine or 1,6-hexanediamine, Examples of charged surface modifications and methods for producing charged surface modifications are given in, for example, See, for example, U.S. Patent No. 6,743,252, which is incorporated herein by reference in its entirety. 3B2 and 7,037,527B2.

[0089] In some cases, the inner and / or outer scaffolding layers of the cell chamber device may be steroids (e.g., dexamethasone, triamcinilone) or immunosuppressants (e.g., thiamin monophosphate) crolimus, sirolimus, everolimus, and / or paclitaxel) In some embodiments, the cell chamber device further comprises a substance that reduces injury-associated inflammation. The inner and / or outer scaffold layers of the implant contain the cytokine CCL22, which Promoting implant tolerance by recruiting Treg cells to the implant's vicinity The substance may be incorporated into the polymer during scaffold production or may be added to the polymer. For example, the material may be coated onto the scaffold after electrospinning. In such an embodiment, the substance can be dissolved in the polymer solution during production. In other embodiments, the nanofibers may be fully or partially formed. The scaffold can be immersed in a solution containing a substance that can be absorbed by the scaffold material. Introducing a net positive or net negative charge into the scaffold fibers as described herein. can promote adhesion of materials loaded onto the scaffold in this manner. , antibiotics, growth factors, etc. can be incorporated into the scaffold material in the manner described above.

[0090] The nanofibrous polymer scaffolds described herein are comprised of random and Additionally or alternatively, the porous material may include any desired pores resulting from the web-like configuration. Pores having shapes and diameters of , nanopores (e.g., having a size of 1 μm or less).

[0091] The pores allow biomolecules secreted by cells within the chamber to exit the scaffold and reach the host tissue or allows blood to pass through while preventing cells from passing in and out of the chamber. For example, the nanopores can be sized to block the flow of the nanopores described herein. biological molecules, e.g., polypeptides, polysaccharides, and / or polynucleotides, as well as organic molecules lipids (e.g., phospholipids, glycolipids, and sterols), vitamins, sugars (e.g., to allow the passage of carbohydrates, disaccharides, oligosaccharides, polysaccharides), and / or amino acids. The pores can be of any size. etc.), peptides, oligopeptides, nucleotides, deoxyribonucleic acid (DNA), ribonucleic acid a size that allows the nucleic acid (RNA) or miRNA to diffuse across the scaffold In certain embodiments, the pore may comprise an antibody, or an antigen-binding portion thereof, scaffolded. It is of such size that it can be passed over.

[0092] In some embodiments, the nanopore is about 250 kDa or less (e.g., about 225 kDa or less). , about 220kDa or less, about 200kDa or less, about 175kDa or less, about 150kDa or less , about 125kDa or less, about 100kDa or less, about 75kDa or less, about 50kDa or less, about allowing the passage of biomolecules with molecular weights of less than 25 kDa, or less than about 10 kDa. In some embodiments, the nanopore is sized to be between 10 kDa and 50 kDa, 0kDa~100kDa, 100kDa~150kDa, 130kDa~165kDa, Biological substances with molecular weights of 150kDa to 200kDa or 200kDa to 250kDa It is sized to allow the passage of molecules.

[0093] The nanopores may also prevent cells from passing through and / or allow cells on one side of the scaffold to penetrate the scaffold. They may be sized to prevent contact with cells on the other side. In embodiments, the nanopores are about 1 μm or less (e.g., about 0.9 μm or less, about 0.8 μm or less). Below, about 0.7μm or less, about 0.6μm or less, about 0.5μm or less, about 0.4μm or less, about 0.3 μm or less, approximately 0.2 μm or less, approximately 0.1 μm or less, or approximately 0.05 μm or less) In certain embodiments, the nanopore has a diameter of 0.5 μm or less. In some embodiments, the nanopores are between 0.1 μm and 0.2 μm, between 0.2 μm and 0.4 μm, 0.3μm~0.5μm, 0.4μm~0.6μm, 0.6μm~0.8μm, or 0 In some embodiments, the nanopore has a diameter of 0.8 μm to 1 μm. In other embodiments, the nanopore has a diameter of between 0.2 μm and 2 μm. In some embodiments, the nanopore is between 1 nm and 10 μm (e.g., between 1 nm and 10 nm, 5 nm m~25nm, 10nm~50nm, 25nm~75nm, 50nm~100nm, 75 nm~125nm, 100nm~200nm, 150nm~250nm, 200nm~3 00nm, 250nm~500nm, 300nm~400nm, 350nm~450nm , 400nm~500nm, 450nm~550nm, 500nm~600nm, 500 nm~1μm, 550nm~650nm, 600nm~700nm, 650nm~750 nm, 700nm~800nm, 750nm~850nm, 800nm~900nm, 9 00nm~1μm, 1μm~2μm, 1μm~5μm, 1μm~10μm, 2μm~4μm m, 2μm~6μm, 2μm~10μm, 4μm~6μm, 4μm~8μm, 4μm~1 0 μm, or 5 μm to 10 μm).

[0094] Antibodies are generally about 10-15 nm in length. Many other peptide therapeutics have similar lengths. For example, Fab fragments have a length of about 9 nm. Therefore, pore sizes of about 10 nm, 15 nm, 20 nm or larger are generally suitable for peptides and The diameter of the nanoparticles is about 10 μm or less (e.g., 9μm or less, 8μm or less, 7μm or less, 6μm or less, 5μm or less, 4μm or less, 3μm or less below, 2 μm or below, or 1 μm or below) are generally used to retain encapsulated cells within the device. The exact size of the pore depends on the size of the encapsulated cell and the size of the secreted biomolecules. It can be adjusted based on the size of the body molecule.

[0095] In some embodiments, the device can be a multi-scaffold device. In addition to the multilayer scaffolds described above, the scaffolds may also include one or more nanofibrous polymer scaffolds. The scaffold includes (i) a first scaffold surrounding a cell chamber, the first scaffold having an outer layer and an inner layer (ii) a first scaffold comprising an outer layer and an inner layer each comprising a nanofibrous polymer; a second scaffold positioned adjacent to the layer and in contact with the cell chamber; The second scaffold may optionally be used for adhesion of cells within the cell chamber. In one embodiment, the second scaffold is a nanofibrous Polymers, including electrospun polymers, for incorporation into the first scaffold Any of the polymers described herein are similarly suitable for incorporation into a secondary scaffold. In an exemplary embodiment, the second scaffold is nPET, nPBT, nPU, or a combination thereof. In an exemplary embodiment, the first The second scaffold comprises nPBT. In some embodiments, the second scaffold comprises pores. Since the scaffold is surrounded by the primary scaffold, the pores can be of any size, e.g., nanoporous or In one embodiment, the second scaffold may be approximately the same size as the first scaffold. In another embodiment, the second scaffold contains pores of approximately the same size as those in the first scaffold. In another embodiment, the second scaffold comprises pores that are larger in size than those in the first scaffold. An exemplary multi-scaffold device containing small pores is shown in Figure ID.

[0096] In some embodiments, the outer (first) scaffold has larger pores than the inner (second) scaffold. For example, in some embodiments, the outer scaffold comprises pores of a size that is larger than the inner chamber bioliquid. Contains pores sized to allow capillary ingrowth for actor perfusion The inner scaffold can be designed to block the passage of cells while allowing the passage of biomolecules. In one embodiment, the outer scaffold comprises pores of a size approximately equal to the diameter of the capillary. Reflects approximately 5-15 μm (e.g., 5-7 μm, 5-10 μm, 5-12 μm, 7-15 The inner scaffold contains pores (e.g., 10-15 μm, 10-15 μm) to hold the cells within the chamber. 1 μm or less (for example, about 0.9 μm or less, about 0.8 μm or less, about 0.7 μm or less, Approx. 0.6 μm or less, Approx. 0.5 μm or less, Approx. 0.4 μm or less, Approx. 0.3 μm or less, Approx. 0.2 1 μm or less, about 0.1 μm or less, or about 0.05 μm or less).

[0097] The scaffold of the cell chamber device is a porous, semi-porous, porous structure positioned between the inner and outer layers. The scaffold may optionally include a porous or non-porous membrane, thereby forming a three-layer scaffold. Membranes are alternatively referred to herein as filters.

[0098] In some embodiments, the membrane can include a nanofibrous polymer. In embodiments, the membrane may comprise nanofibrous polybutylene terephthalate. The fibrous membrane results from a random web-like arrangement of nanofibers, as described above for the scaffold. It may contain pores.

[0099] In other embodiments, the membrane is not nanofibrous. For example, the membrane is made of nanofibers of a predetermined size. Non-nanofibrillar membranes may include, for example, PET, PB, or other materials. PU, or any of the polymers described herein. It can be composed of, without limitation, virtually any biocompatible polymer. The non-nanofibrillar membrane may comprise one or more of the following polymers: mixed cellulose; Cellulose acetate (MCE), cellulose acetate, coated cellulose acetate, hydrophilic PTFE, hydrophobic PTFE, nylon, or polycarbonate. In some embodiments, The electrospun fibers can coat one or both sides of the membrane. , solid (non-nanofibrous) films containing pores of defined sizes, e.g., nPE It can be coated with electrospun fibers such as T-PBT or nPU. In one embodiment, the scaffold comprises electrospun nPET-PBT on one side and electrospun nPET-PBT on the other side. A nanopore-studded solid (non-nanofiber) coated with electrospun nPU Nanopore-studded solid membranes include non-nanofiberous polymer sheets ( For example, PET, PBT, PU, ​​or other suitable polymers, or combinations thereof. In an exemplary embodiment, the solid film comprises PET. In an exemplary embodiment, the solid membrane comprises PU. Incorporation of a non-nanofibrillar membrane into the scaffold The nanofibrous material may function as a barrier to block the passage of cells, otherwise In such an embodiment, the membrane may be sized to block the passage of cells. Scaffolds containing a solid (non-nanofibrous) membrane as the outer surface can be embedded in the implant. At the implantation site, electrospun fibers may cause inflammation in the host subject. Coating a solid (non-nanofibrous) membrane with The inflammatory response to the scaffold was significantly reduced compared to a scaffold consisting of a solid membrane without coating. It can be reduced.

[0100] Membrane nanopores allow biomolecules, e.g., proteins, to cross the membrane while blocking their passage through cells. In one embodiment, the membrane pores may be sized to allow passage of approximately 250 kDa. or less (e.g., about 225 kDa or less, about 220 kDa or less, about 200 kDa or less, about 17 5kDa or less, about 150kDa or less, about 125kDa or less, about 100kDa or less, about 75 kDa or less, about 50 kDa or less, about 25 kDa or less, or about 10 kDa or less) In some embodiments, the membrane pores are sized to allow passage of biomolecules having , 10kDa~50kDa, 50kDa~100kDa, 100kDa~150kDa, Biological substances with molecular weights of 150kDa to 200kDa or 200kDa to 250kDa The membrane pores are sized to allow the passage of molecules. They also prevent the passage of cells, and and / or a surface that prevents cells on one side of the scaffold from contacting cells on the other side of the scaffold. Thus, in some embodiments, the membrane may be about 1 μm or smaller (e.g., Approximately 0.9μm or less, approximately 0.8μm or less, approximately 0.7μm or less, approximately 0.6μm or less, approximately 0.5 μm or less, approx. 0.4 μm or less, approx. 0.3 μm or less, approx. 0.2 μm or less, approx. 0.1 μm or less In certain embodiments, the nanopores have a diameter of about 0.05 μm or less. In some embodiments, the membrane nanopore has a diameter of 0.5 μm or less. 0.1μm~0.4μm, 0.4μm~0.6μm, 0.6μm~0.8μm, or 0 In some embodiments, the membrane nanopores have a diameter of 0.1 to 3 μm. In another embodiment, the membrane nanopore has a diameter of 0.2 to 2 μm. In some embodiments, the membrane nanopores are between 1 nm and 10 μm (e.g., between 1 nm and 10 nm, 5 nm m~25nm, 10nm~50nm, 25nm~75nm, 50nm~100nm, 75 nm~125nm, 100nm~200nm, 150nm~250nm, 200nm~3 00nm, 250nm~500nm, 300nm~400nm, 350nm~450nm , 400nm~500nm, 450nm~550nm, 500nm~600nm, 500 nm~1μm, 550nm~650nm, 600nm~700nm, 650nm~750 nm, 700nm~800nm, 750nm~850nm, 800nm~900nm, 9 00nm~1μm, 1μm~2μm, 1μm~5μm, 1μm~10μm, 2μm~4μm m, 2μm~6μm, 2μm~10μm, 4μm~6μm, 4μm~8μm, 4μm~1 In some embodiments, the membrane pores have a diameter of nanometers (0 μm, or 5 μm to 10 μm). The pores are approximately the same size as the pores in the outer and / or inner layers of the fibrous polymer scaffold. In embodiments, the membrane pores are smaller than the pores in the outer and / or inner layers of the nanofibrous polymer scaffold. In other embodiments, the membrane pores are formed by micropores in the outer and / or inner layers of the nanofibrous polymer scaffold. larger than the hole.

[0101] In some embodiments, one or more electrospun polymers can be porous, semi-porous, or or non-porous membrane to form a three-layer material, which can be used to , some embodiments of the cell chamber devices described herein can be fabricated. In such an embodiment, a polyester film or membrane (e.g., a polyester film or membrane of a predetermined size) is used. For example, about 3 mm to about 300 mm, for example, 3 mm to 10 mm, 10 mm to 25 mm, 25 mm~50mm, 50mm~75mm, 75mm~100mm, 100mm~125mm , 125mm~150mm, 150mm~175mm, 175mm~200mm, 200 mm~225mm, 225mm~250mm, 250mm~275mm, or 275mm m, or 300 mm) is the mandrel of the electrospinning unit or the grounded The electrospinning unit can then be loaded onto the collector. One or more electrospun polymers on one or both sides of a polyester film or membrane -Operate according to standard operating procedures for coating (e.g., PET and PBT) In certain embodiments, the electrospun material can be applied to one side of the membrane. The other side of the membrane is coated with nPET-PBT and / or nPU. For example, the device may include a film or membrane (e.g., a PET membrane) coated with a Apply an nPET / PBT layer to the membrane, and an nPU layer on top of the nPET / PBT layer on one side of the membrane. In another embodiment, the device comprises a scaffold comprising a film or membrane. A film or a film with an nPET / PBT layer applied on one side of the film and an nPU layer applied on the other side of the film. The scaffold may include a membrane. Optionally, an adhesive may be applied to one or both sides of the membrane. In some embodiments, electrosparging can be used to help bond the layers together. The adhesive is applied between the membrane and the nanofibrous polymer layer deposited thereon. In embodiments, the electrospun adhesive is applied to the same electrospun layer as one or more of the other layers. The material may include a fibrous material.

[0102] Similar to the inner or outer layer of a scaffold, the nanoporous film or membrane may be used to connect the scaffold to the host tissue. allowing the passage of biomolecules secreted by cells into the tissue or blood, and and / or allow the passage of biomolecules (e.g., nutrients) from the host into the chamber to The membrane may contain nanopores sized to accommodate cells seeded in the device. For example, The membrane nanopore can be configured to contain any of the biomolecules described herein, e.g., polypeptides, polysaccharides, and polynucleotides, and lipids (e.g., phospholipids, glycolipids, and sterols), Chemical messengers (e.g., hormones and neurotransmitters), vitamins, sugars (e.g., charcoal), hydrates, disaccharides, oligosaccharides, polysaccharides), amino acids, peptides, oligopeptides, polypeptides peptides, proteins, nucleotides, deoxyribonucleic acid (DNA), or ribonucleic acid (R The size may be such that it allows the passage of organic molecules such as NAs. In this configuration, the membrane nanopore allows the passage of a protein, e.g., an antibody, or antigen-binding portion thereof. It is sized to make it possible.

[0103] In some embodiments, the membrane nanopore is about 250 kDa or less (e.g., about 225 kDa or less). Lower, about 220kDa or less, about 200kDa or less, about 175kDa or less, about 150kDa or less Lower, about 125kDa or less, about 100kDa or less, about 75kDa or less, about 50kDa or less, Allows passage of biomolecules with molecular weights of approximately 25 kDa or less, or approximately 10 kDa or less In some embodiments, the membrane nanopores are sized to accommodate nanopores between 10 kDa and 50 kDa. , 50kDa~100kDa, 100kDa~150kDa, 130~165kDa, 1 Biocomponents with molecular weights of 50 kDa to 200 kDa or 200 kDa to 250 kDa The size is such that it allows the passage of a child.

[0104] In an exemplary embodiment, the cell chamber device comprises a multi-layer scaffold surrounding the cell chamber. The multilayer scaffold comprises nanofibrous polyethylene terephthalate and polybutylene terephthalate. an outer layer including a polyurethane foam and an inner layer including a nanofibrous polyurethane, nm to 10um (e.g., 1nm to 10nm, 5nm to 25nm, 10nm to 50nm, 25nm~75nm, 50nm~100nm, 75nm~125nm, 100nm~20 0nm, 150nm~250nm, 200nm~300nm, 250nm~500nm, 300nm~400nm, 350nm~450nm, 400nm~500nm, 450n m~550nm, 500nm~600nm, 500nm~1μm, 550nm~650n m, 600nm~700nm, 650nm~750nm, 700nm~800nm, 75 0nm~850nm, 800nm~900nm, 900nm~1μm, 1μm~2μm, 1μm~5μm, 1μm~10μm, 2μm~4μm, 2μm~6μm, 2μm~10μm m, 4μm-6μm, 4μm-8μm, 4μm-10μm, or 5μm-10μm) In another exemplary embodiment, the cell chamber device includes a nanopore having a diameter. The scaffold comprises a triple-layered scaffold surrounding the cystic chamber, the triple-layered scaffold comprising nanofibrous polyethylene terephthalate. an outer layer comprising polyethylene terephthalate and polybutylene terephthalate, and an inner layer comprising nanofibrous polyurethane. and a nanofibrous polybutylene terephthalate layer positioned between the inner layer and the outer layer. and a film, and the outer layer, inner layer, and film are 1 nm to 10 μm (for example, 1 nm to 10 nm, 5nm~25nm, 10nm~50nm, 25nm~75nm, 50nm~100nm, 75nm~125nm, 100nm~200nm, 150nm~250nm, 200nm ~300nm, 250nm~500nm, 300nm~400nm, 350nm~450 nm, 400nm~500nm, 450nm~550nm, 500nm~600nm, 5 00nm~1μm, 550nm~650nm, 600nm~700nm, 650nm~7 50nm, 700nm~800nm, 750nm~850nm, 800nm~900nm , 900nm~1μm, 1μm~2μm, 1μm~5μm, 1μm~10μm, 2μm~ 4μm, 2μm~6μm, 2μm~10μm, 4μm~6μm, 4μm~8μm, 4μm Contain nanopores with diameters of 10 μm to 10 μm, or 5 μm to 10 μm.

[0105] In some embodiments, one or more layers of the scaffold comprise pores having a diameter of 1 μm or less.

[0106] The cell chamber devices described herein can be used for administration (e.g., minimally invasive procedures) to a host subject. any size suitable for administration via a surgical procedure, e.g., laparoscopic or endoscopic surgery) The size and shape may be modified as needed to accommodate the device. Fitting the device to the implantation site, containing the desired number of cells that can be loaded, and / or or a dosage of a therapeutic biomolecule secreted by cells within the device to a recipient subject. Regulate (e.g., measured by mg of biomolecule secreted from the device per day) can be).

[0107] In some embodiments, the cell chamber contains up to about 1 x 10 11 cells (e.g., up to approximately 1×10 11 cells, up to approximately 1 x 10 10 cells, up to approximately 1 x 10 9 cells, maximum Approximately 1×10 8 cells, up to approximately 1 x 10 7 cells, up to approximately 1 x 10 6 cells, up to approximately 1×10 5 cells, up to approximately 1 x 10 4 cells, or up to approximately 1 x 10 3 cells) In some embodiments, the cell chamber is sized to accommodate up to 1 x 10 7 pieces In some embodiments, the cell chamber is sized to accommodate up to 1× 10 11 The device is sized to accommodate 100 cells. The number of cells loaded into the device is Based on the desired dose of biomolecule to be delivered and the amount of biomolecule produced by the cells, For example, a dose of 10 μg per day may be required, and the If the cells produce 1 pg / cell / day, the chamber contains 1 x 10 7 to accommodate individual cells It can be designed to.

[0108] The cell chamber device may be any suitable device suitable for stable implantation at the desired delivery site of the biomolecule. Thus, in various embodiments, the device can be substantially The shape may be cylindrical, flat, disc-shaped, patch-shaped, oval, star-shaped, tubular, or spherical. In some embodiments, the cell chamber device is planar or substantially planar. In embodiments, the cell chamber device is planar or substantially planar and may be rectangular, square, or shaped as or approximately shaped as a triangle, circle, pentagon, hexagon, heptagon, or octagon In other embodiments, the device is not planar. For example, in some embodiments In this case, the device is shaped as or approximately shaped as a sphere, cylinder, rod, cube, etc. In some embodiments, the shape of the cell chamber may be arranged in a spiral or It can be folded to maximize surface area.

[0109] In certain embodiments, the cell chamber device is rectangular. In embodiments, the rectangular cell chamber device may be about 15 cm or less, about 12 cm or less, about 1 0cm or less, about 8cm or less, about 6cm or less, about 4cm or less, about 2cm or less, about 1cm or less , about 0.1 cm (100 mm) or less in length, or about 0.01 cm (10 mm) or less in length In some embodiments, the rectangular cell chamber device can be approximately 15 cm Below, about 12cm or less, about 10cm or less, about 8cm or less, about 6cm or less, about 4cm or less, Approximately 2 cm or less, approximately 1 cm or less, approximately 0.1 cm (100 mm) or less, or approximately 0.01 cm In some embodiments, the cell chamber device has a width of about 5 to 10 mm. In some embodiments, the cell chamber has a width of 10 cm and a length of about 10-15 cm. The device has a width of about 3-5 cm and a length of about 5-10 cm. The cell chamber device has a width of about 1 to 3 cm and a length of about 3 to 5 cm. In embodiments, the cell chamber device has a width of about 0.01 to 1 cm and a length of about 1 to 3 cm. In some embodiments, the cell chamber device has a thickness of about 0.01 cm, about 0.1 cm, approx. 1cm, approx. 2cm, approx. 3cm, approx. 4cm, approx. 5cm, approx. 6cm, approx. 7cm, approx. 8 cm, approx. 9cm, approx. 10cm, approx. 11cm, approx. 12cm, approx. 13cm, approx. 14cm, and a first dimension of about 15 cm, and a second dimension of about 0.01 cm, about 0.1 cm, about 1 cm, about 2 cm, Approximately 3cm, approximately 4cm, approximately 5cm, approximately 6cm, approximately 7cm, approximately 8cm, approximately 9cm, approximately 10cm , a second dimension of approximately 11 cm, approximately 12 cm, approximately 13 cm, approximately 14 cm, or approximately 15 cm In one particular embodiment, the cell chamber device has a width of about 8 cm and a thickness of about 10 cm. In other embodiments, the cell chamber device has a width of about 5 cm and a length of about 9 In other embodiments, the cell chamber device has a width of about 3 cm and a length of about In other embodiments, the cell chamber device has a width of about 1 cm and a length of about 5 cm. It is about 3 cm long.

[0110] In some embodiments, the device is about 250 cm 2 For example, about 225 cm 2 below , about 220cm 2 Below, approximately 200cm 2 Below, approximately 175cm 2 Below, approximately 150cm 2 below , about 125cm 2Below, approximately 100cm 2 Below, approximately 75cm 2 Below, approximately 50cm 2 Below, approx. 25cm 2 Less than or equal to 10cm 2 It has the following surface area:

[0111] In some embodiments, the device is about 5 cm 3 For example, about 4.5 cm 3 Below, approx. 4cm 3 Below, approximately 3.5cm 3 Below, approximately 3cm 3 Below, approximately 2.5cm 3 Below, approximately 2cm 3 Below, approximately 1.5cm 3 Below, approximately 1cm 3 Less than or equal to 0.5 cm 3 It has the following volume: .

[0112] The device may be of any suitable thickness to accommodate the desired implantation site. In some embodiments, the device is substantially planar and has a second smallest dimension, e.g., a For example, 20% or less of width, 15% or less, 10% or less, 9% or less, 8% or less, 7% or less, 6% including a total thickness that is not more than 5%, not more than 4%, not more than 3%, not more than 2%, or not more than 1%. In some embodiments, the device is about 1 mm or less, 0.9 mm or less, 0.8 mm or less, 0 .7mm or less, 0.6mm or less, 0.5mm or less, 0.4mm or less, 0.3mm or less, 0 In some embodiments, the device comprises a total thickness of 0.2 mm or less, or 0.1 mm or less. , about 250 μm or less (e.g., about 225 μm or less, about 220 μm or less, about 200 μm or less) , about 175 μm or less, about 150 μm or less, about 125 μm or less, about 100 μm or less, about 75 The thickness of the film may be less than about 50 μm, less than about 25 μm, or less than about 10 μm. In some embodiments, the device is between 10 μm and 50 μm, between 10 μm and 100 μm, between 50 μm m~100μm, 50μm~200μm, 100μm~150μm, 100μm~250 μm, 100μm~500μm, 150μm~200μm, 200μm~250μm, 2 50μm~300μm, 250μm~500μm, 300μm~350μm, 350μm ~400μm, 400μm~450μm, 450μm~500μm, 500μm~550 μm, 500μm~1mm, 550μm~600μm, 600μm~650μm, 650 μm~700μm, 700μm~750μm, 750μm~800μm, 750μm~1 mm, 800μm~850μm, 850μm~900μm, 900μm~950μm, or a total thickness of 950 μm to 1 mm. In certain embodiments, the device Includes total thickness of less than μm.

[0113] The cell chamber devices disclosed herein optionally include a method for transferring cells into the cell chamber. It may also include a loading port to facilitate loading. Such a loading port may be used to The loading port can be located on the outer surface of the cell chamber device facing the outside. An opening (e.g., for inserting a cell loading or injection device) and a surface on the inside of the cell chamber Deposit cells into the cell chamber via an internal opening (e.g., a cell loading or injection device) The loading port can include a loading port for loading cells into the cell chamber. After cell loading, the cells may be sealed to prevent leakage of the ultrasonic waves from the cell chamber. The opening in the chamber can be sealed by various methods, such as welding. Other examples of sealing processes known in the art include heat staking (e.g., cold staking). taking or heat staking), swaging, spin welding, hot plate welding These include welding, vibration welding, or laser welding.

[0114] III.Cells The devices disclosed herein contain cells in a cell chamber surrounded by a multilayer scaffold. Thus, in some embodiments, the device may optionally include The cells can be loaded via a cell loading port or an opening in the bilayer scaffold. The cells can be loaded into the cell chamber via, for example, The cell chamber can then be sealed using the method. The inner layer of the bilayer scaffold may optionally be coated with a layer of cellulose to allow the cells to adhere to the inner layer of the scaffold. So you will be contacted.

[0115] In some embodiments, the cell line selected for use in the cell chamber device can have one or more of the following characteristics: (1) the cells can be grown under harsh conditions; (2) the cells can be adapted to produce (e.g., secrete) the desired therapeutic biomolecule; (3) cells have a relatively long life or shelf life (e.g., (4) If the subject is a human, the cells may be encapsulated. (5) cells that can be of human origin to increase compatibility between the cells and the host; can exhibit high viability in the device to ensure long-term delivery (e.g., (e.g., greater than 80% viability in the device for greater than one month in vivo); (6) encapsulation The cells are capable of delivering an effective amount of a useful biological product, (7) the cells are have a low level of host immune response to ensure longevity of the graft, and and / or (8) the cells are non-tumorous to provide additional safety in the event of device leakage. It could be.

[0116] In some embodiments, mammalian proteins known in the art for the production of mammalian proteins are used. Mammalian cells may be suitable for use in the cell chamber device. Chinese Hamster Ovary (CHO) cells, as well as, for example, mouse myeloma (NS0), baby hamster kidney (B various other mammalian cells, such as human embryonic kidney (HEK-293), human embryonic kidney (HEK-293), and human retinal cells Cell lines obtained from the source are approved by regulatory agencies for the manufacture of biopharmaceuticals, including therapeutic antibodies. Examples of mammalian host cells include CHO, BHK, and HEK293C. OS, PC12, HiB5, RN33b, C2C12, HepG2, and ARPE-19 Examples include cells.

[0117] In certain embodiments, the cells are human cells, including recombinant cells of human origin. In embodiments, the cells are human retinal pigment epithelial (RPE) cells, or cells derived therefrom. In an exemplary embodiment, the cells comprise ARPE-19 cells. ARPE-19 cells strains (e.g., Dunn et al., 62 Exp. Eye Res. 155-69 (1 996), Dunn et al,39 Invest.Ophthalmol.Vis .Sci.2744-9(1998), Finnemann et al.,94 Pr oc.Natl.Acad.Sci.USA 12932-7(1997), Handa et al.,66 Exp.Eye.411-9(1998), Holtkamp et al., 112 Clin.Exp.Immunol.34-43(1998), Maidji et al.,70 J.Virol.8402-10(1996), US No. 6,361,771 (see U.S. Patent No. 6,361,771) is a cell chamber as described herein. It has many characteristics of a platform cell line for use with the device. The E-19 cell line was purchased from the American Type Culture Collection ARPE-19 cells are available from the American College of Cardiology (ATCC No. CRL-2302). CRAL is a specific marker for RPE cells. ARPE-19 cells express BP and RPE-65. They form stable monolayers, which The ARPE-19 cell line exhibits morphological and functional polarity during implantation into a host subject. and are genetically engineered to secrete biomolecules of therapeutic interest. It can be genetically modified, has a relatively long life span, and is of human origin. The ARPE-19 cells implanted in the device had good in vivo viability and were compatible with human hosts. In another embodiment, the cells elicit a non-significant immune response and are not tumorigenic. are human hepatocytes. In another embodiment, the cells are human pancreatic islet cells.

[0118] The cells may be transformed or non-transformed. Furthermore, the cells may be in suspension or In some embodiments, the cells are contact-inhibited cells, e.g., For example, immortalized contact-inhibited human cells (e.g., the hTERT immortalized cell line (Evercyte, V The cells are known in the art for recombinant protein production. For example, mammalian cell lines that may be used in the devices described herein include In some embodiments, the SV40-transformed monkey kidney CVI strain (COS- 7, ATCC™ CRL1651), baby hamster kidney cells (BHK, ATCC (TMCCL10), mouse Sertoli cells (TM4, Mather, Biol. Rep rod., 23:243 (1980)), monkey kidney cells (CVI-76, ATCC (trademark) ) CCL70), African green monkey kidney cells (VERO-76, ATCC™ CRL -1587), canine kidney cells (MDCK, ATCC™ CCL34), buffalo Rat liver cells (BRL3A, ATCC.RTM.CRL1442), mouse breast cancer cells ( MMT060562, ATCCCV CCL51), rat hepatocytes (HTC, MI.54, Baumann et al., J. Cell Biol., 85:1 (1980)) 3T3 cells, 293T cells (Pear, WS, et al., Proc. Natl. Acad.Sci.USA.,90:8392-8396(1993)), NS0 cells ( Sato et al.Tissue Culture Association,24 :1223(1988)), SP2 / 0(Sato et al.J.Exp.Med. , 165:1761 (1987)), TR-1 cells (Mather et al., An nals NYAcad.Sci.,383:44(1982)), and hybrids In some embodiments, the cell line is a human embryonic kidney line 293S (G raham et al., J. Gen. Virolo., 36:59 (1977)), Human cervical cancer cells (HELA, ATCC CCL2), human lung cells (W138, A TCC™ CCL75), human liver cells (Hep G2.HB8065), hTER T immortalized cell line (Evercyte, Vienna, AT), or human retinal cells In some embodiments, the cell line is a human cell line. A carcinogenic cell line that has been modified or treated to make it safe for use in the device In some embodiments, the cell chamber device can be used to culture stem cells, such as induced pluripotent stem cells ( human stem cells, such as iPS cells, embryonic stem cells (ES), or mesenchymal stem cells (MSC), or differentiated cells derived from such stem cells.

[0119] In some cases, the cells loaded into the cell chamber devices of the present disclosure are, e.g., , genetically engineered cells that have been engineered to produce, e.g., secrete, a biomolecule of interest, e.g., For example, recombinant cells may be used as biomolecules that can be secreted by cells of the device. Examples include, but are not limited to, polypeptides, polysaccharides, and polynucleotides. and lipids (e.g., phospholipids, glycolipids, and sterols), chemical messengers (e.g., neurotransmitters and hormones such as insulin), vitamins, sugars (e.g., carbohydrates), , disaccharides, oligosaccharides, polysaccharides), amino acids, peptides, oligopeptides, polypeptides, proteins proteins, nucleotides, deoxyribonucleic acid (DNA), or ribonucleic acid (RNA) Other secreted biomolecules include exosomes, lipid polymers, and These may include those assembled, packaged, and secreted as viral particles. In embodiments, the cells produce a therapeutic biomolecule, as described herein.

[0120] In some embodiments, the cells in the device are capable of expressing one or more proteins or peptides, e.g. For example, one or more recombinant proteins or peptides, e.g., one or more therapeutic proteins. For example, cells within the device may be engineered to secrete antibodies, proteins, or peptides. or antigen-binding fragments thereof, growth factors, hormones (e.g., insulin), cytokines , coagulation factors (e.g., factor VIII or factor IX, or variants thereof, e.g., Recombinate, Kogenate, Refacto, Advate, Al prolix, BeneFIX, Rixubis, Ixinity, Idelvion etc. or a combination thereof. The protein may, in some cases, be a recombinant protein or peptide. obtain.

[0121] In some embodiments, the cells in the device express one protein or peptide, e.g. The recombinant or therapeutic protein or peptide can be secreted. In embodiments, the cells in the device express two or more proteins, e.g., 2, 3, 4, 5, 6 , 7, 8, 9, 10 or more recombinant or therapeutic proteins or peptides. In some embodiments, the device can secrete multiple proteins or peptides. In other embodiments, the cells can be loaded with a single cell type, e.g., a single cell line. The device may comprise two or more, each of which secretes one or more proteins or peptides. For example, loading 2, 3, 4, 5, 6, 7, 8, 9, 10 or more cell types, e.g., cell lines For example, a cell chamber device can be used to target two secreted proteins. In one embodiment, the device is used to provide two recombinant proteins or In another embodiment, the device can be loaded with cells that secrete the peptide. Loading two cell lines, each secreting a single recombinant protein or peptide can be done.

[0122] In some embodiments, the cells in the device are induced by fibroblast growth factor (FGF), epidermal growth factor (EGF), or fibroblast growth factor (FGF). factor (EGF), platelet-derived growth factor (PDGF), insulin-like growth factor (IGF), Transforming growth factor (TGF), vascular endothelial growth factor (VEGF), hepatic growth factor (LGF) , bone morphogenetic proteins (BMPs), colony-stimulating factors (CSFs), hepatocyte growth factors (HGFs) ), or nerve growth factor (NGF), or combinations thereof. The cells can secrete one or more growth factors, which are not specified.

[0123] In some embodiments, the cells within the device are selected from the group consisting of bone morphogenetic proteins (BMPs), Erythrocytes, and the like. Epoietin (EPO), granulocyte colony-stimulating factor (G-CSF), granulocyte macrophage Colony-stimulating factor (GM-CSF), interferon alpha, interferon b interleukin 2 (IL-2), interleukin 11 (IL-11), or interferon gamma, or combinations thereof secrete more than one cytokine.

[0124] In some embodiments, the cells in the device are capable of expressing insulin, estrogen, progestogens, and thyroxine (as levothyroxine), or steroids, or combinations thereof secrete one or more hormones, including but not limited to:

[0125] In some embodiments, the cells in the device secrete one or more enzymes. The cells within the chair are able to secrete enzymes that provide enzyme replacement therapy (ERT) to the subject. In some embodiments, the cells express hexosaminidase A, alpha-galactosidase A , glucocerebrosidase, arylsulfatase A, galactocerebrosidase, and Lysosomal storage disease deficiencies, including but not limited to, sphingomyelinase and lysosomal enzyme deficiencies, In some embodiments, the cells are capable of producing complementing enzymes. Also known as mucopolysaccharidosis type II (MPSII), which includes, but is not limited to, In some embodiments, the cells are capable of producing an enzyme that complements Hunter syndrome. Metachromatic leukodystrophy, including but not limited to arylsulfatase A ( In some embodiments, the cells are capable of producing enzymes that complement the Laroni including, but not limited to, enzymes It can produce enzymes that complement mucopolysaccharidosis type I (MPSI).

[0126] In some embodiments, the cells in the device contain one or more antibodies, or antigen-binding portions thereof. The antibodies, or antigen-binding portions thereof, described herein can be secreted in their entirety. long antibodies, bispecific antibodies, dual variable domain antibodies, multi-chain or single-chain antibodies, and / or Fab, Fab', (Fab')2, Fv, scFv (single-chain Fv), surrobody ( antibodies, including synthetic light chain constructs), single domain antibodies, camelized antibodies, etc. They may also be in the form of antigen-binding fragments that specifically bind to extracellular molecules that are not specifically targeted to the antibody. For example, IgA (e.g., IgA1 or IgA2), IgD, IgE, IgG (e.g., any isotype, including IgG1, IgG2, IgG3, or IgG4, or IgM In some embodiments, the Cells include Fab, F(ab')2, scFv, tandem scFv, diabodies, minibodies, In some embodiments, the antibody fragment is selected from the group consisting of a single domain antibody, a single domain antibody, and a single domain antibody. In this state, the cells secrete full-length antibodies.

[0127] In some embodiments, the cells secrete the chimeric antibody, or an antigen-binding portion thereof. The term "antibody" refers to an antibody whose variable region sequences are derived from one species and whose constant region sequences are derived from another species. Antibodies derived from, for example, mouse antibodies whose variable region sequences are derived from murine antibodies and whose constant region sequences are derived from human antibodies The term "antibody" is intended to refer to an antibody that

[0128] In some embodiments, the cells secrete a humanized antibody, or an antigen-binding portion thereof. "Humanized" forms of human (e.g., rodent) antibodies contain minimal sequence derived from the non-human antibody. For the most part, humanized antibodies are chimeric antibodies that are engineered to target a human "recipient." Residues from the hypervariable regions of the antibody are selected from mouse, rat, and other from the hypervariable regions of a "donor" antibody from a non-human species such as mouse, rabbit, or non-human primate In the recipient antibody, the residues of the human immunoglobulin (recipient antibody) are replaced by those of the following: In some cases, framework region (FR) residues of human antibodies are substituted with corresponding non-human residues. Furthermore, in some cases, humanized antibodies may be modified by the recipient These modifications can include residues that are not found in either the recipient or donor antibody. The performance of the antibody can be further improved. In some embodiments, the humanized antibody comprises a hypervariable C All or substantially all of the DR loops correspond to hypervariable loops of a non-human antibody, and all of the FR loops or at least one, typically two, possible FRs substantially all of which are human antibody sequences. Humanized antibodies can optionally contain substantially all of the antibody constant region. The antibody will also contain at least a portion of the constant region (Fc), typically a human antibody. For further details, see, for example, Jones et al., Nature 321:522 -525(1986), Riechmann et al., Nature332:32 3-329 (1988), and Presta, Curr. Op. Struct. Biol. .2:593-596(1992).

[0129] In some embodiments, the cells secrete human antibodies, or antigen-binding portions thereof. The term "antibody" as used herein refers to both the framework and CDR regions. It also refers to an antibody having variable regions derived from human germline immunoglobulin sequences. Where the antibody contains constant regions, the constant regions also may be derived from human germline immunoglobulin sequences. The human antibodies of the invention are derived from human germline immunoglobulin sequences (e.g., in vitro by random or site-directed mutagenesis in vitro or by somatic mutation in vivo. However, the amino acid sequence may contain amino acid residues not encoded by the The term "human antibody" as used herein refers to an antibody derived from another mammalian species, such as a mouse. an antibody in which CDR sequences derived from the germline of is not intended to include.

[0130] In some embodiments, the cells secrete monoclonal antibodies. The term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies. That is, the individual antibodies that make up the population are identical in sequence and specificity. may contain naturally occurring mutations or may arise during the production of a monoclonal antibody preparation. This excludes mutant antibodies that may be mutated and such mutants are generally present in small amounts. Polyclonal antibodies, typically containing different antibodies directed against different determinants (epitopes) In contrast to monoclonal antibody preparations, each monoclonal antibody binds to a single antigen. Thus, the modifier "monoclonal" refers to a substantially homogeneous antibody. The antibody is characterized as being obtained from a population of antibodies, and the antibody is characterized by the presence of a specific antibody. It is not to be construed as requiring isolation. For example, Monoclonal antibodies are produced using hybridoma, recombinant DNA, and phage display methods. and transgenic animals containing all or part of the human immunoglobulin locus The ion channel may be derived using a variety of techniques, including but not limited to, methods utilizing Such methods and other exemplary methods for producing monoclonal antibodies are described in the art. These techniques are known in the art and are described herein.

[0131] If the cells in the cell chamber secrete antibodies, or antigen-binding portions thereof, The antibody or antigen-binding portion thereof can specifically bind to any antigen of interest. In embodiments, the antibody, or antigen-binding portion thereof, binds to (i) a tumor-associated antigen, (ii) a cell surface antigen, receptors, (iii) CD proteins and their ligands, e.g., CD3, CD4, C D8, CD19, CD20, CD22, CD25, CD32, CD33, CD34, CD 40, CD44, CD47, CD54, CD59, CD70, CD74, CD79a(C CD79a), and CD79P (CD79b), (iv) members of the ErbB receptor family (v) a receptor, e.g., an EGF receptor, a HER2, a HER3, or a HER4 receptor; cell adhesion molecules, e.g. LFA-1, Mac1, p150,95, VLA-4, ICAM- 1, VCAM, and αv / β3 containing either alpha or beta subunits Integrin (e.g., anti-CD11a, anti-CD18, or anti-CD11b antibodies), or ( vi) growth factors, e.g., PDGF, FGF, VEGF, IgE, blood group antigens, flk2 / flt3 receptor, obesity (OB) receptor, mpl receptor, CTLA4, protein C, protein B R3, c-met, tissue factor, etc. In some embodiments, the antibody, or antigen-binding portion thereof, is selected from the group consisting of 4-1BB, 5T4, ACVR2B, ADA M-9, alpha-V integrin, AMHRII, AXL, BAFF, BAFF-R, Basigin, BCMA, C242 antigen, c-Met, CA9, CA-125, CanAg , CCR2, CCR4, CCR5, CD2, CD3, CD3 epsilon, CD3E, CD 4, CD5, CD6, CD11, CD11a, CD15, CD18, CD19, CD20 , CD22, CD23, CD25, CD27, CD28, CD30, CD33, CD37 , CD38, CD40, CD40L, CD41, CD44v6, CD45, CD49b, CD51, CD52, CD54, CD56, CD70, CD74, CD79B, CD80 , CD97B, CD98, CD99, CD117, CD123, CD125, CD134 , CD137, CD138, CD147, CD152, CD154, CD163, CD1 66, CD184, CD200, CD205, CD221, CD248, CD276, C D278, CD279, CD319, CD352, CDH-6, CEA, CEA-CAM 4, CEA-CAM5, CEA-Cide, CEA-related antigen, CEACAM1, CEAC AM6, CFC1B, claudin-18 isoform 2, CLDN6, and CLDN18. 2, CSF1R, CTLA4, CXCR4, dendritic cell-associated lectin 2, DLK1, DLL 3, DLL4, DR5, EFNA4, EGFR, EGFR extracellular domain III, EGF Rviii, endoglin, endothelin receptor ETB, ENPP3, EpCAM, Ep hA, EPHA3, ephrin receptor A3, ephrin A4, episialin, ER-alpha 36, ERBB1, ERBB2, ERBB3, FCGRT, FGFR, FGFR2, Bronectin extra domain-B, FLT3, folate receptor, folate receptor 1, folate receptor receptor alpha, FOLHI, Frizzled receptor, FXYD5, ganglioside GD 3. GCC, GCGR, GCL, GD2 ganglioside, GD3 ganglioside, globoside H, glypican 3, GMCSF receptor α chain, GPC2, GPNMB, granulocyte antigen, GUC Y2C, H-ferritin, hepatitis B surface antigen, HER1, HER2, HER2 / neu, HER3, HGFR, HLA-DR, human scatter factor receptor kinase, ICAM-1, IC OS, IgE receptor, IGF1R, IL6R, IL31RA, IL3 receptor, IL-4R α, IL-6R, IL-12 / 23, IL-17 receptor, integrin α4, integrin Integrin α4β7, integrin α5β1, integrin αIIbβ3, integrin αvβA , integrin β7, interferon receptor, interferon α / β receptor, ITG A2, ITGB2, KAAG-1, KIR2D, L-selectin, CD62L, LAG3 , LAMP1, Le(y), LFA-1, LINGO-1, LIV-1, LRRC15, LY6E, LYPD3, MCAM, mesothelin, MS4A1, MSLN, MST1R, M T1-MMP, MTX3, MTX5, MUC1, mucin 16, mucosal addressin cell adhesion component MAdCAM, myelin-associated glycoprotein, NCA-90, nectin-4, NGN A ganglioside, NKA, NKG2A, Notch3, Notch1, Notch receptor body, NRP1, OFP, NaPi2b, OX-40, P-cadherin, P. aeruginosa nosa antigen, PCDC1, PD-1, PD-L1, PDCD1, PDGF-Rα, PD GFRA, sodium phosphate cotransporter, phosphatidylserine, platelet-derived growth factor receptor Body beta, PRLR, PSMA, PTK7, RGMA, RHD, Rhesus factor, Rhesus Ror1, Ror2, RSVFR, SAIL, SDC1, selectin P, SL AMF7, SLC34A2, SLC44A4, SLeA, SLITRK5, SLITRK 6, Soluble IL-6, SOST, SSTR2, STEAP1, STn, T cell receptor, T ACSTD2, TAG-72, TEM1, TF, TIGIT, TIM-1, TM4SF1 , TNF-α, TNFR superfamily member 4, TNFRSF17, TRAIL -R1, TRAIL-R2, TROP2, TWEAK receptor, VEGFA, VEGFR2 , VEGFR-1, VSIR, or VWF. Antigens that can be targeted by antibodies, or antigen-binding fragments thereof Other examples include, for example, Chen and F, which are incorporated herein by reference. lies.Nature reviews immunology.13.4(2013 ):227.

[0132] In some embodiments, the antibody secreted by the cells in the device is 3F8 (GD2 gas). abciximab (ReoPro; binds to CD41), abituzumab (binding to CD51), alemtuzumab (Lemtrada, Campath; CD52 abrilumab (binds to integrin α4β7), adalimumab (Humira ; binds to TNF-α), adecatumumab (binds to EpCAM), alacizumab pegol (V binds to EGFR2), alemtuzumab (Lemtrada, Campath; binds to CD52) Altumomab pentetate (Hybri-ceaker; CEA-bound), Tuximab (conjugated to mesothelin), anatumomab mafenatox (conjugated to TAG-72) , anetumab ravtansine (bound to MSLN), anifrolumab (interferon α / β receptor), apolizumab (binding to HLA-DR), apolizumab ixadotin ( FGFR2), arcitumomab (CEA), acelizumab (L-selectin or CD62L), and atezolizumab (Tecentriq; binds to PD-L1) , atrolimumab (binding to rhesus factor), avelumab (Bavencio; PD-L 1), abicixizumab (binding to DLL4; VEGFA), azintuxizumab Dotin (binds to CD319), basiliximab (Simulect; binds to CD25), Bavituximab (binding to phosphatidylserine), BCD-100 (binding to PD-1), Vectamumomab (LymphoScan; binds to CD22), belantamab mafodotin (binding to BCMA), belimumab (Benlysta; binding to BAFF), bemarituzumab (binding to FGFR2), benralizumab (Fasenra; binding to CD125), Sanlimab (binding to ICAM-1), besilesomab (Scintimun; CEA-associated antibody) gen), bimagrumab (binding to ACVR2B), bivatuzumab mertansine (binding to CD4 4 v6), bleselumab (binding to CD40), blinatumomab (Blincyt o; binds to CD19), brosozumab (binds to SOST), brentuximab vedotin ( Adcentris (binding to CD30), brontizumab (binding to Notch1), Lodalumab (Siliq; binds to IL-17 receptor), cabilalizumab (binds to CSF1R) (binding to CD25), Camerelizumab (binding to PD-1) , carotuximab (binding to endoglin), catumaxomab (Removab; EpCA M / CD3), cantuzumab ravtansine (binding to MUC1), caplacizumab ( Cablivi (binding to VWF), cedelizumab (binding to CD4), semipilimab (Li btayo (binding to PCDC1), cetrelimab (binding to PD-1), certolizumab ( TNF-α binding), sergituzumab amnalikine (CEA binding), cetuximab ( Erbitux (binding to EGFR), Sivisatamab (binding to CEACAM5), Sirmuts IgE-1 receptor (binding to CD221), cixutumumab (binding to ROR1), Lenoliximab (binding to CD4), clivatuzumab tetraxetan (hPAM4-Cid e; binds to MUC1), codrituzumab (binds to glypican 3), coltuximab cisplatin (binding to CD19), conatumumab (binding to TRAIL-R2), crizanlisma (binding to selectin P), clotidumab (binding to GCGR), dacetuzumab (binding to CD4 0), daclizumab (Zenabata; binds to CD25), dalotuzumab (IGF -1 receptor (binding to CD221), dapirolizumab pegol (binding to CD154; binding to CD40L) daratumumab (Darzalex; binds to CD38), demcizumab (DLL4) denituzumab mafodotin (binding to CD19), deptuzumab mafodotin drozitumab (binding to DR5), DS-8201 (binding to HER2) binding to HER3), deligotuzumab (ERBB3; binds to HER3), dinutuximab (Unitux) in; binds to GD2 ganglioside), dupilumab (binds to IL-4Rα), durba lumab (Imfinzi; binds to PD-L1), duvortuximab (CD19; CD3 E), ecloneximab (binding to GD3 ganglioside), edrecolomab (binding to Ep CAM), elezanumab (RGMA), elgemtumab (ERBB3, HE R3), elotuzumab (binding to SLAMF7), emactuzumab (binding to CSF1R enapotamab vedotin (binding to AXL), enavatuzumab (binding to TWEAK receptor) enlimonomab pegol (binding to ICAM-1; CD54) Enobrituzumab (binding to CD276), Enoticumab (binding to DLL4), Epstein-Barr virus Latuzumab (binds to CD22), erlizumab (binds to ITGB2; CD18), Ertu Makizumab (Rexomun; HER2 / neu; binds to CD3), etaracizumab (A bergin; integrin α v β3 binding), etigilimab ) (binding to TIGIT), etrolizumab (binding to integrin β7), exvivir (binding to hepatitis B surface antigen), fanolesomab (NeutroSpec; binding to CD15) Faralimomab (binding to interferon receptors), farletuzumab (binding to folate receptors) FBTA05 (Lymphomun; binds to CD20), Fugachipotsuma fgatipotuzumab (binding to MUC1), fivatuzumab (binding to ephrin receptor) Figitumumab (binding to IGF-1 receptor; CD221), Flotezumab Izumab (binding to IL3 receptor), foralumab (binding to CD3 epsilon), Futuximab galiximab (binding to EGFR), galiximab (binding to CD80), gancotamab (binding to HER2 / n eu), ganitumab (IGF-1 receptor; binds to CD221), gabirimomab (C D147; binds to basigin), gemtuzumab ozogamicin (Mylotarg; CD3 3), glenbatumumab vedotin (binding to GPNMB), golimumab (binding to Simpo ni; binds to TNF-α), gomiliximab (CD23; binds to IgE receptor), iana Ibalizumab (binding to BAFF-R), Ibalizumab (binding to CD4), I BI308 (binding to PD-1), ibritumomab tiuxetan (binding to CD20), Rucumab (binding to VEGFR-1), ifavotuzumab (binding to EPHA3), igovoma IMAB362 (Indimacis-125; binds to CA-125), IMAB362 (CLDN1 8.2), imaprelimab (binding to MCAM), inlacumab (binding to selectin P) binding), indatuximab ravtansine (binding to SDC1), iradatuzumab vedotin ( CD97B), imgatuzumab (binding to EGFR), indusatumab vedotin ( GUCY2C), inebilizumab (binding to CD19), infliximab (Rem icade (binding to TNF-α), intetumumab (binding to CD51), inolimomab ( binds to CD25), inotuzumab ozogamicin (Besponsa; binds to CD22) , ipilimumab (Yervoy; binds to CD152), Iomab-B (binds to CD45) , iratumumab (binding to CD30), isatuximab (binding to CD38), iscalimab (binds to CD40), istiratumab (binds to IGF1R; CD221), itolizumab (Alzumab; binds to CD6), Keliximab (binds to CD4), Laprituximab Emtansine (binding to EGFR), labetuzumab (CEA-Cide; binding to CEA), Rifastuzumab vedotin (binding to sodium phosphate cotransporter), remaresomab (NCA -90 (binding to granulocyte antigen), lembervimab (binding to hepatitis B surface antigen), leronrimab ribivirumab (binding to CCR5), lexatumumab (binding to TRAIL-R2), Hepatitis C surface antigen (binding to hepatitis C surface antigen), roncaximab tesirin (binding to CD19), rosatuximab Mabvedotin (binds to EGFR; ERBB1; HER1), rilotomab tetraxetan (binding to CD37), lintuzumab (binding to CD33), lirilumab (binding to KIR2D ), lorvotuzumab mertansine (binding to CD56), lucatumumab (binding to CD40) , lulizumab pegol (binding to CD28), lumiliximab (binding to CD23; IgE receptor) ), lumletuzumab (ERBB3; binds to HER3), rupartumab amadotin (LY PD3), mapatumumab (TRAIL-R1), margetuximab (HER 2), maslimomab (binding to T cell receptor), mavrilimumab (binding to GMCSF receptor) α chain), matuzumab (binding to EGFR), milatuzumab (binding to CD74), Retumomab (binding to TAG-72), mirvetuximab soravtansine (binding to folate receptor agonist) fa), mitumomab (binding to GD3 ganglioside), morolimumab (rhesus monkey factor), modotuximab (binding to EGFR extracellular domain III), mogamulizumab (binding to CCR4), monalizumab (binding to NKG2A), mosunetuzumab (binding to CD3E ;MS4A1; binds to CD20), moxetumomab pasudotox (binds to CD22), mu Romonab-CD3 (binds to CD3), nacolomabutafenatox (binds to C242 antigen) , naptumomab estafenatox (binding to 5T4), naratuximab emtansine ( CD37), nalunatumab (MST1R), natalizumab (Tysabri ; binds to integrin α4), naxitamab (binds to c-Met), necitumumab (EG FR), nemolizumab (IL31RA), nimotuzumab (Theracim Theraloc (binding to EGFR), nirsevimab (binding to RSVFR), nivolumab PD-1 binding, obinutuzumab (CD20 binding), ocaratu- zumab (CD20 binding) ocrelizumab (binding to CD20), odulimomab (binding to LFA-1; CD11 a), ofatumumab (binding to CD20), olatumab (binding to PDGF-Rα) , omburtamab (binding to CD276), onartuzumab (human scatter factor receptor kinase Ontuxizumab (binding to TEM1), Onvatilimab (binding to VSIR), Opicinumab (binding to LINGO-1), oportozumab monatox (binding to EpCAM) Oregovomab (binding to CA-125), otelixizumab (binding to CD3), Reltuzumab (binding to CD37), oxelumab (binding to OX-40), panitumumab ( EGFR), vancomab (binding to tumor-specific glycosylation of MUC1), patitus ERBB3 (binding to HER3), PDR001 (binding to PD-1), pembrolizumab Keytruda (binding to PD-1), pemtumomab (Theragyn; binding to MUC1 pertuzumab (Omnitarg; binds to HER2 / neu), pidilisma (binding to PD-1), pinatuzumab vedotin (binding to CD22), prosalizumab (binding to CD22) CR2), pogalizumab (binding to TNFR superfamily member 4), pola Tuzumab vedotin (binding to CD79B), prilidimab (binding to CD4), PRO140 (binding to CCR5), ramucirumab (Cyramza; binding to VEGFR2), Mab (binding to CD40), leratolimab (binding to LAG3), linucumab (platelet-derived augmentation rituximab (binding to CD20), rituzimab (binding to growth factor receptor beta), hera; Rituzan; binds to CD20), lobatumumab (IGF-1 receptor; CD 221), racotumomab (Vaxira; binds to NGNA ganglioside), rad Retumab (binding to fibronectin extra domain-B), refanezumab (myelination) Leukemia-associated glycoprotein), loredumab (binding to RHD), robelizumab (binding to Leukemia-associated glycoprotein), Arrest (binds to CD11; CD18), rozanolixizumab (binds to FCGRT) , ruplizumab (Antova; binds to CD154; CD40L), SA237 (IL- 6R), sacituzumab govitecan (binding to TROP-2), samalizumab (binding to CD2 00), sarilumab vedotin (binding to LRRC15), sarilumab (Kevza) ra; binds to IL-6R), satralizumab (binds to IL6 receptor), satumomab pende tido (binding to TAG-72), seribantumab (binding to ERBB3; HER3), Setol Sumab (binding to SOST), SGN-CD19A (binding to CD19), SHP647 (binding to adhesion) membrane addressin (binding to cell adhesion molecules), siltuximab (Sylvant; soluble IL-6 , which binds to IL-6R), siplizumab (which binds to CD2), siltratumab ( tumab), vedotin (binding to SLITRK6), sontuzumab (binding to episialin) , sofituzumab vedotin (binding to CA-125), solitomab (binding to EpCAM), Spartalizumab (PDCD1; binds to CD279), suresomab (NCA-90; granules) (binding to leukocyte antibody), sputumab (binding to RSVFR), tabalumab (binding to BAFF), Tadocizumab (integrin α IIbβ3), talacuzumab (binding to CD123 ), taplitumomab paptox (binding to CD19), talectumab (binding to Notch receptor ), tavolimab (binding to CD134), telisotuzumab vedotin (binding to HGFR) ), teneliximab (binding to CD40), tepoditamab (binding to dendritic cell-associated lectin 2) Teprotumomab (IGF-1 receptor; binds to CD221), tetulomab (CD37 TGN1412 (binding to CD28), tiburizumab (binding to BAFF), Tuzumab (binding to TRAIL-R2), timigituzumab (binding to HER2), tiragotumab (binding to TIGIT), tislelizumab (binding to PCDC1; CD279), tocilizumab Mab (Actemra; RoActemra; binds to IL-6 receptor), tomzotuximab (EGFR; binds to HER1), toralizumab (CD154; binds to CD40L), Situmomab (Bexxar; binds to CD20), tobetumab (binds to PDGFRA), Rustuzumab (Herceptin; binds to HER2 / neu), trastuzumab emta cisplatin (Kadcyla; binds to HER2 / neu), tregalizumab (binds to CD4) , tremelimumab (binding to CTLA4), TRBS07 (binding to GD2 ganglioside) , tucotuzumab-cell molleukin (binding to EpCAM), ublituximab (binding to MS4A1) binds to CXCR4; binds to CD184), urocupramab (4-1BB; binds to CD137), ustekinumab (Stellera; binds to IL-12 / 23), Utomilumab (4-1BB; binds to CD137), vadastuximab butaririn (CD33 (binding to Frizzled receptors), vanalimab (binding to CD40), vantictumab (binding to Frizzled receptors) (binding to VEGFR2), valisacumab (binding to VEGFR2), varlilumab (binding to CD27), telizumab (ITGA2; binds to CD49b), vedolizumab (Entyvio; binds to intracellular signaling) tRNA α4β7), veltuzumab (binding to CD20), besencumab (binding to NRP1) binds to CD3), visilizumab (Nuvion; binds to CD3), bovalilizumab (binds to IL6R) volociximab (binding to integrin α5β1), bonlerolizumab (binding to CD134 ), vopratelimab (CD278; binds to ICOS), XMAB-5574 (CD 19), zalutumumab (HuMax-EGFr; binds to EGFR), zanolimumab (HuMax-CD4; binds to CD4), zatuximab (binds to HER1), Xenoctu Mab (ERBB3; binds to HER3), Dillalimumab (CD147; binds to basigin), Zolbetuximab (binding to claudin-18 isoform 2) or zolimonad ritox (binding to CD5), or antigen-binding portions thereof.

[0133] Other antibodies that may be secreted by cells within the device include anetumab (anti-mesothelin binding), aortumab (binding to FGFR2), azintuximab (binding to SLAMF7) , belantamab (binding to TNFRSF17), bivatuzumab (binding to CD44v6), Lentuximab (binding to CD30), Camidanlumab (binding to CD25), Cantuzumab (binding to CanAg), cantuzumab (binding to CanAg), clivatuzumab (MUC1 cofetuzumab (binding to PTK7), coltuximab (binding to CD19), Nintuzumab (binding to CD19), depatuximab (binding to EGFR), enapotamab ( AXL), enfortumab (binding to nectin-4), epratuzumab (binding to CD22) Gemtuzumab (binding to CD33), Glembatumumab (binding to GPNMB), Hemtuzumab Rutuzumab (binding to HER2), iradatuzumab (binding to CD79B), indatuximab (binding to CD138), indutuzumab (binding to GCC), inotuzumab (binding to CD2 2), labetuzumab (binding to CEA-CAM4), ladiratuzumab (binding to LIV-1 ), laprituximab (binding to EGFR), rifastuzumab (binding to SLC34A2) loncastuzumab (binding to CD19), lorvotuzumab (binding to CD56), Satuximab (binding to EGFR), Rupartumab (binding to LYPD3), Iratumumab ( binds to CD30), milatuzumab (binds to CD74), mirvetuximab (binds to PSMA) naratuximab (binding to CD37), pinatuzumab (binding to CD22), polatuzumab umab (binding to CD79B), rovalpituzumab (binding to DLL3), sacituzumab (TA CSTD2), samtrotamab (binding to LRRC15), siltratumab (SLT RK6), sofituzumab (binding to mucin 16), telisotuzumab (binding to c-Met) tisotumab (binding to TF), trastuzumab (binding to ERBB2), vadastuki citabine (binding to CD33), bundurumab (binding to STEAP1), or bolus Antigen-binding fragments include, but are not limited to, tuzumab (which binds to CD70), or antigen-binding portions thereof. Not determined.

[0134] In some embodiments, the cells in the cell chamber device specifically bind to α4β7. In certain embodiments, the cells secrete antibodies, or antigen-binding portions thereof. In some embodiments, the cells in the device secrete a mab, or an antigen-binding portion thereof. Anti-α4β7 antibodies such as vedolizumab, or binding molecules containing the antigen-binding region of vedolizumab Vedolizumab is a commercial product of mammalian host cells engineered to stably express the Name: ENTYVIO® (Millennium Pharmaceuticals) Vedolizumab is also known as a medicament for targeting α4β7 integrins, e.g. , which specifically binds to the α4β7 complex and is a mucosal addressin and cell adhesion component of the α4β7 integrin. Blocks interactions with MAdCAM-1 and fibronectin, and transendothelial Lymphocytes, e.g., CD4, CD8, and memory T lymphocytes, in the inflamed gastrointestinal parenchyma Vedolizumab is a humanized monoclonal antibody that inhibits the migration of α4β1 and It does not bind to or inhibit the function of αEβ7 integrins, but acts as a vascular cell adhesion It does not antagonize the interaction with vascular endothelial cell adhesion molecule-1 (VCAM-1).

[0135] α4β7 integrin is a distinct subset of memory T lymphocytes that preferentially migrate to the gastrointestinal tract. MAdCAM-1 is expressed primarily on the surface of intestinal epithelial cells and is expressed on T lymphocytes. It plays an important role in lymphocyte homing to intestinal lymphoid tissues. Interaction with MAdCAM-1 is a hallmark of ulcerative colitis and Crohn's disease It has been implicated as an important factor in mucosal inflammation, including chronic inflammation. Colitis and ulcerative colitis, pouchitis (including, for example, chronic pouchitis), graft-versus-host disease, celiac disease Chronic kidney disease, HIV, primary sclerosing cholangitis, and MAdCAM or fibronectin Inflammatory bowel disease, including mucosal inflammation from lymphocyte trafficking after adhesion to α4β7 ligands It can be used to treat.

[0136] In one embodiment, the cells in the cell chamber device comprise a heavy chain variable region of SEQ ID NO: 1, and and / or secrete an antibody comprising the light chain variable region of SEQ ID NO: 5, or an antigen-binding portion thereof. In embodiments, the cells in the cell chamber device comprise a heavy chain CDR1 of SEQ ID NO: 2, a heavy chain CDR2 of SEQ ID NO: 3, a heavy chain CDR3 of SEQ ID NO: 4, a heavy chain CDR4 of SEQ ID NO: 5, a heavy chain CDR5 of SEQ ID NO: 6, a heavy chain CDR6 of SEQ ID NO: 7, a heavy chain CDR7 of SEQ ID NO: 8, a heavy chain CDR8 of SEQ ID NO 3, and a heavy chain CDR2 of SEQ ID NO: 4, and / or a light chain CDR3 of SEQ ID NO: 6 R1, an antibody comprising a light chain CDR2 of SEQ ID NO: 7, and a light chain CDR3 of SEQ ID NO: 8, or In one embodiment, the cells in the cell chamber device secrete the antigen-binding portion. A heavy chain comprising the amino acid sequence of SEQ ID NO:9 and / or a light chain comprising the amino acid sequence of SEQ ID NO:10 Vedolizumab and its derivatives are also known to be effective in treating rheumatoid arthritis. The columns are based on U.S. Patent Publication Nos. 2014 / 0341885 and 2014 / 037 and No. 7251, each of which is expressly incorporated herein by reference in its entirety. The cells in the devices herein contain the binding sites described above and in the enclosed sequence listing. The antibody can be engineered to secrete antibodies containing a region, e.g., a CDR or variable region. .

[0137] In certain embodiments, the cells in the cell chamber device contain the amino acid sequence of SEQ ID NO:1. an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 5, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 6; In some embodiments, the cells in the cell chamber device secrete an antigen-binding portion thereof. The cell comprises CDR1 of SEQ ID NO: 2, CDR2 of SEQ ID NO: 3, and CDR3 of SEQ ID NO: 4. Heavy chain variable region, and CDR1 of SEQ ID NO: 6, CDR2 of SEQ ID NO: 7, and CDR2 of SEQ ID NO: 8 Some antibodies secrete antibodies containing a light chain variable region comprising the CDR3 of In embodiments, the cells in the cell chamber device contain a heavy chain comprising the amino acid sequence of SEQ ID NO:9. and an antibody comprising a light chain comprising the amino acid sequence of SEQ ID NO: 10, or an antigen-binding portion thereof. In certain embodiments, the cells in the cell chamber device secrete vedolizumab or or secrete the antigen-binding portion thereof.

[0138] In one embodiment, the cells in the cell chamber device are stimulated with tumor necrosis factor alpha (TNFα In one embodiment, the cell secretes an antibody, or an antigen-binding portion thereof, that specifically binds to the The cells in the chamber device contain the heavy chain variable region of SEQ ID NO: 22 and / or the heavy chain variable region of SEQ ID NO: 23 In one embodiment, the cell secretes an antibody, or an antigen-binding portion thereof, comprising the light chain variable region of The cells in the chamber device contained heavy chain CDR1 of SEQ ID NO: 24, heavy chain CDR2 of SEQ ID NO: 25 2, and the heavy chain CDR3 of SEQ ID NO: 26, and / or the light chain CDR1 of SEQ ID NO: 27, An antibody comprising a light chain CDR2 of SEQ ID NO: 28 and a light chain CDR3 of SEQ ID NO: 29, or an antibody thereof In one embodiment, the cells in the cell chamber device secrete the original binding moiety. 0, and / or a light chain comprising the amino acid sequence of SEQ ID NO: 31. The cells in the device herein secrete antibodies, or antigen-binding portions thereof, containing: Antibodies comprising the binding regions, e.g., CDRs or variable regions, set forth above and in the enclosed sequence listings. can be engineered to secrete

[0139] In one embodiment, the cells in the cell chamber device are treated with interleukin-12 (IL-1 2) and secrete an antibody, or an antigen-binding portion thereof, that specifically binds to the antigen. The cells in the cell chamber device contain the heavy chain variable region of SEQ ID NO: 32 and / or the heavy chain variable region of SEQ ID NO: 3 In one embodiment, the cells secrete antibodies, or antigen-binding portions thereof, comprising the light chain variable region of each of the three antibodies. The cells in the cell chamber device were incubated with heavy chain CDR1 of SEQ ID NO: 34, heavy chain CDR2 of SEQ ID NO: 35 R2, and a heavy chain CDR3 of SEQ ID NO: 36, and / or a light chain CDR1 of SEQ ID NO: 37, an antibody comprising a light chain CDR2 of SEQ ID NO: 38 and a light chain CDR3 of SEQ ID NO: 39, or In one embodiment, the cells in the cell chamber device secrete the antigen-binding moiety. a heavy chain comprising the amino acid sequence of SEQ ID NO: 40 and / or a light chain comprising the amino acid sequence of SEQ ID NO: 41 The cells in the device herein secrete antibodies, or antigen-binding portions thereof, containing the antibodies. and antibodies comprising the binding regions, e.g., CDRs or variable regions, set forth above and in the enclosed sequence listing. The body can be engineered to secrete

[0140] In some embodiments, the cell chamber device comprises a chamber coated with an antibody, or an antigen-binding portion thereof. For example, a gene encoding an antibody, or antigen-binding portion thereof, as described above may be used. one or more nucleic acid molecules operably linked to a promoter and / or other elements necessary for In one embodiment, the cell chamber device comprises a cell containing an antibody heavy chain and / or a is a nucleic acid sequence comprising one or more nucleic acids encoding an antibody light chain or a portion thereof. and recombinant host cells containing the above expression vector(s).

[0141] For example, in some embodiments, the cell chamber device may contain 3F8 (GD2 ganglioside). abciximab (ReoPro; binds to CD41), abituzumab (CD5 1), alemtuzumab (Lemtrada, Campath; binds to CD52), Abrilumab (binding to integrin α4β7), adalimumab (Humira; TNF- α), adecatumumab (binding to EpCAM), alacizumab pegol (binding to VEGFR2 binds to CD52), alemtuzumab (Lemtrada, Campath; binds to CD52), Rutumomab pentetate (Hybri-ceaker; CEA-bound), amatuximab (bound to mesothelin), anatumomab mafenatox (bound to TAG-72), anetu Mabravtansine (binding to MSLN), Anifrolumab (interferon α / β receptor apolizumab (binding to HLA-DR), apolizumab ixadotin (binding to FGFR 2), arcitumomab (binding to CEA), acelizumab (binding to L-selectin or C D62L), atezolizumab (Tecentriq; binds to PD-L1), Limumab (binding to rhesus monkey factor), avelumab (Bavencio; binding to PD-L1) ), abicixizumab (binding to DLL4; VEGFA), azintuxizumab vedotin ( binds to CD319), basiliximab (Simulect; binds to CD25), Cimab (binding to phosphatidylserine), BCD-100 (binding to PD-1), Vectam Momab (LymphoScan; binds to CD22), belantamab mafodotin (BCM A), belimumab (Benlysta; bound to BAFF), bemarituzumab (FG FR2), benralizumab (Fasenra; binds to CD125), Belsanrima (binding to ICAM-1), besilesomab (scintimun; binding to CEA-related antigen) ), bimagrumab (binding to ACVR2B), bivatuzumab mertansine (CD44 v6 Blincyto (CD40 binding), bleselumab (CD40 binding), blinatumomab (CD 19), brosozumab (SOST), brentuximab vedotin (Adce ntris (binds to CD30), brontizumab (binds to Notch1), brodalumab Siliq (binding to IL-17 receptor), cabilalizumab (binding to CSF1R), Midanlumabtesirin (binding to CD25), Camrelizumab (binding to PD-1), Caroline Ximab (binding to endoglin), catumaxomab (Removab; EpCAM / CD 3), cantuzumab ravtansine (binding to MUC1), caplacizumab (binding to Cabl ivi; binds to VWF), cedelizumab (binds to CD4), semipilimab (Libtay o; binds to PCDC1), cetrelimab (binds to PD-1), certolizumab (binds to TNF- α), sergituzumab amnalikine (CEA binding), cetuximab (Erbi tux; binds to EGFR), sibisatamab (binds to CEACAM5), cirumutuzumab ( ROR1), cixutumumab (binding to IGF-1 receptor; CD221), and clenol Cimab (binding to CD4), clivatuzumab tetraxetan (hPAM4-Cide; MU C1), codrituzumab (binding to glypican 3), coltuximab ravtansine ( CD19), conatumumab (TRAIL-R2), crizanlizumab (Celeron Clottedumab (binding to GCGR), Dacetuzumab (binding to CD40) ), daclizumab (Zenabata; binds to CD25), dalotuzumab (IGF-1 receptor dapirolizumab pegol (CD154; binds to CD40L), dapirolizumab pegol (CD154; binds to CD40L), Ratumumab (Darzalex; binds to CD38), Demcizumab (binds to DLL4), Denintuzumab mafodotin (binding to CD19), depatuxizumab mafodotin (EG FR), dorozizumab (DR5), DS-8201 (HER2), Rigotuzumab (ERBB3; binds to HER3), dinutuximab (Unituxin; G D2 ganglioside), dupilumab (IL-4Rα), durvalumab ( Imfinzi (binding to PD-L1), Duvortuximab (binding to CD19; binding to CD3E) ), ecloneximab (binding to GD3 ganglioside), edrecolomab (binding to EpCAM) elezanumab (binding to RGMA), elgemtumab (binding to ERBB3, HER3) Elotuzumab (binding to SLAMF7), emactuzumab (binding to CSF1R), Napotamab vedotin (binding to AXL), enavatuzumab (binding to TWEAK receptor), Enlimonomab pegol (binding to ICAM-1; CD54), enoblitzumab (binding to CD276 DLL4), enoticumab (binding to DLL4), epratuzumab (binding to CD22), rituximab (ITGB2; binds to CD18), ertumakizumab (Rexomun; HER 2 / neu; binds to CD3), etaracizumab (Abergin; integrin α v β3 Etigilimab (binding to TIGIT), etrolizumab (binding to integrin β7) binding), exbivirumab (binding to hepatitis B surface antigen), fanolesomab (Neutro Spec; binds to CD15), faralimomab (binds to interferon receptor), Luretuzumab (binding to folate receptor 1), FBTA05 (lymphomun; binding to CD20) fgatipotuzumab (binding to MUC1), Matuzumab (binding to ephrin receptor A3), fizitumumab (binding to IGF-1 receptor; CD221), flotetuzumab (binding to IL3 receptor), foralumab (CD3 epsilon futuximab (binding to EGFR), galiximab (binding to CD80), Cotamab (binding to HER2 / neu), ganitumab (binding to IGF-1 receptor; CD22 1), gabirimomab (binding to CD147; basigin), gemtuzumab ozogamicin (M ylotarg (binds to CD33), glenbatumumab vedotin (binds to GPNMB), Golimumab (Simponi; binds to TNF-α), golimumab (CD23; IgE receptor), ianalumab (binding to BAFF-R), ibalizumab (Trogarz o; binds to CD4), IBI308 (binds to PD-1), ibritumomab tiuxetan ( CD20), icrucumab (binding to VEGFR-1), ifavotuzumab (EPH A3), igovomab (Indimacis-125; binds to CA-125), IM AB362 (binding to CLDN18.2), imaprelimab (binding to MCAM), Incl. Coumab (binding to selectin P), indatuximab ravtansine (binding to SDC1), Radatuzumab vedotin (binding to CD97B), imgatuzumab (binding to EGFR), Dusatumab vedotin (binding to GUCY2C), inebilizumab (binding to CD19), Infliximab (Remicade; binds to TNF-α), intetumumab (binds to CD51) binding), inolimomab (binding to CD25), inotuzumab ozogamicin (Bespon sa; binds to CD22), ipilimumab (Yervoy; binds to CD152), iomab -B (binding to CD45), iratumumab (binding to CD30), isatuximab (binding to CD38 iscalimab (binding to CD40), istiratumab (binding to IGF1R; CD 221), Itolizumab (Alzumab; binds to CD6), Keliximab (binds to CD4) combination), laprituximab emtansine (binding to EGFR), labetuzumab (CEA-Ci de; binds to CEA), rifastuzumab vedotin (binds to sodium phosphate cotransporter) , remaresomab (NCA-90; binds to granulocyte antigen), lembervimab (hepatitis B surface antigen) leronlimab (binding to CCR5), lexatumumab (binding to TRAIL-R2) combined), ribivirumab (binding to hepatitis B surface antigen), loncastuximab tesirin (CD1 9), rosatuxizumab vedotin (binding to EGFR; ERBB1; HER1), Lonsurfaxtetraxetan (binding to CD37), lintuzumab (binding to CD33), Lumab (binding to KIR2D), lorvotuzumab mertansine (binding to CD56), Lucat lumalizumab (binding to CD40), lulizumab pegol (binding to CD28), lumiliximab (binding to CD40) D23 (binding to IgE receptor), ramletuzumab (ERBB3 (binding to HER3)), Paltumab amadocin (binding to LYPD3), mapatumumab (binding to TRAIL-R1) , margetuximab (binding to HER2), maslimomab (binding to T cell receptors), mabri Limumab (GMCSF receptor α chain), matuzumab (binding to EGFR), milatuzumab (C D74), minletumomab (TAG-72), mirvetuximab soravtan folate receptor α binding, mitumomab (binding to GD3 ganglioside), morolimbap (binding to rhesus factor), modotuximab (binding to EGFR extracellular domain III) , mogamulizumab (binding to CCR4), monalizumab (binding to NKG2A), mosnetuz mab (binding to CD3E; MS4A1; CD20), moxetumomab passudotox (CD 22), muromonab-CD3 (binding to CD3), nacolomabutafenatox (binding to C2 42 antigen), estafenatox (binding to 5T4), naratuximab Mutansin (binding to CD37), nalunatumab (binding to MST1R), natalizumab (T ysabri (binding to integrin α4), naxitamab (binding to c-Met), necits tumab (binding to EGFR), nemolizumab (binding to IL31RA), nimotuzumab (Th eracim; Theraloc; binds to EGFR), nirsevimab (binds to RSVFR) ), nivolumab (binding to PD-1), obinutuzumab (binding to CD20), okara ocrelizumab (binding to CD20), odulimomab (binding to LFA- 1; binds to CD11a), ofatumumab (binds to CD20), olaratumab (binds to PDGF -Rα), omburtamab (binding to CD276), onartuzumab (binding to human scatter factor receptor kinase), ontuxizumab (binding to TEM1), onvatilimab (VS IR), opicinumab (binding to LINGO-1), oportozumab monatox ( EpCAM), oregovomab (binding to CA-125), otelixizumab (binding to CD3 otlertuzumab (binding to CD37), oxelumab (binding to OX-40), Panitumumab (binding to EGFR), vancomab (binding to tumor-specific glycosylation of MUC1) ), patizumab (ERBB3; binds to HER3), PDR001 (binds to PD-1), Pembrolizumab (Keytruda; binds to PD-1), pemtumomab (Therag yn; binds to MUC1), pertuzumab (Omnitarg; binds to HER2 / neu) , pidilizumab (binding to PD-1), pinatuzumab vedotin (binding to CD22), pro Zalizumab (binding to CCR2), Pogalizumab (TNFR superfamily member 4 polatuzumab vedotin (binding to CD79B), priligimab (binding to CD4) , PRO140 (binding to CCR5), ramucirumab (Cyramza; binding to VEGFR2) ), ravagalimab (binding to CD40), raletimab (binding to LAG3), linucumab ( Platelet-derived growth factor receptor beta), rituximab (binding to CD20), (MabThera; Rituzan; binds to CD20), lobatumumab (IGF-1 receptor; binds to CD221), Racotumomab (Vaxira, binds to NGNA ganglioside radletumab (binding to fibronectin extra domain-B), Refanez Mab (binding to myelin-associated glycoprotein), loredumab (binding to RHD), rovelizumab LeukArrest (binding to CD11 and CD18), rozanolixizumab (FCG RT), ruplizumab (Antova; binds to CD154; binds to CD40L), SA2 37 (binding to IL-6R), sacituzumab govitecan (binding to TROP-2), Samariz Mab (binding to CD200), samrotamab vedotin (binding to LRRC15), sarilumab (Kevzara; binds to IL-6R), satralizumab (binds to IL6 receptor), Momab pendetide (binding to TAG-72), seribantumab (binding to ERBB3; HER3) Setrusumab (binds to SOST), SGN-CD19A (binds to CD19), SH P647 (binding to mucosal addressin cell adhesion molecule), silmutuzumab (Sylvant; Soluble IL-6 (binding to IL-6R), siplizumab (binding to CD2), siltratumab Vedotin (binding to SLITRK6), sontuzumab (binding to episialin), Sofituz Mabvedotin (binding to CA-125), solitomab (binding to EpCAM), Spartaliz Mab (PDCD1; binds to CD279), Sulesomab (NCA-90; binds to granulocyte antibody) ), sputumab (binding to RSVFR), tabalumab (binding to BAFF), tadocizumab (Integrin α IIb β3), talacuzumab (binding to CD123), taprit Momabpaptox (binding to CD19), tarexuzumab (binding to Notch receptor), and tabo limab (binding to CD134), telisotuzumab vedotin (binding to HGFR), tenelixirix Mab (binding to CD40), tepoditamab (binding to dendritic cell-associated lectin 2), teproz Momab (IGF-1 receptor; binds to CD221), tetulomab (binds to CD37), TG N1412 (binding to CD28), tiburizumab (binding to BAFF), tigatuzumab (TR AIL-R2), timigituzumab (binding to HER2), tiragotumab (TIGIT ), tislelizumab (binding to PCDC1; CD279), tocilizumab (binding to Acte mra; RoActemra; binds to the IL-6 receptor), tomzotuximab (EGFR; HER1), toralizumab (CD154; CD40L), tositumomab (B exxar (binds to CD20), tobetumab (binds to PDGFRA), trastuzumab ( Herceptin (binding to HER2 / neu), trastuzumab emtansine (Kad cyla (binding to HER2 / neu), tregalizumab (binding to CD4), tremelimumab (binding to CTLA4), TRBS07 (binding to GD2 ganglioside), tucotuzumab Cermoleukin (binding to EpCAM), ublituximab (binding to MS4A1), urokinase Plumab (CXCR4; binds to CD184), urelumab (4-1BB; binds to CD137) ), ustekinumab (Stellera; binds to IL-12 / 23), utomilumab ( 4-1BB (binding to CD137), vadastuximab butarilin (binding to CD33), limumab (binding to CD40), vanticutumab (binding to Frizzled receptors), Sacumab (binding to VEGFR2), varlilumab (binding to CD27), batelizumab (I TGA2 (binding to CD49b), vedolizumab (Entyvio; binding to integrin α4β7 veltuzumab (binding to CD20), besencumab (binding to NRP1), visilii Ibizumab (Nuvion; binds to CD3), bovalilizumab (binds to IL6R), boroxicam Shima (binding to integrin α5β1), Bonlerolizumab (binding to CD134), Laterimab (binding to CD278; ICOS), XMAB-5574 (binding to CD19), Zalutumumab (HuMax-EGFr; binds to EGFR), zanolimumab (HuMax- CD4 (binding to CD4), Zatuximab (binding to HER1), Xenoctuzumab (ERBB 3; binds to HER3), diralimumab (CD147; binds to basigin), zolbetuximab (binding to claudin-18 isoform 2), or zolimomab aritox (C D5), or one or more nucleic acid molecules encoding antigen-binding portions thereof Preferably, the nucleic acid molecule is nucleated by the cells present in the chamber. A promoter and / or other components necessary for the expression and secretion of the protein encoded by the is operably linked to the regulatory element of

[0142] In some embodiments, the cells in the cell chamber device disclosed herein are α4β 7, or one or more nucleic acids encoding an antibody that specifically binds to, or an antigen-binding portion thereof. In certain embodiments, the cells are vedolizumab or an antigen-binding portion of vedolizumab. The nucleic acid sequence encoding the light chain variable region is SEQ ID NO: 1. The nucleic acid sequence encoding the heavy chain variable region is set forth in SEQ ID NO: 12. The full-length nucleic acid sequence encoding the light chain of dolizumab is shown as SEQ ID NO: 13. The full-length nucleic acid sequence encoding the heavy chain of vedolizumab is shown as SEQ ID NO: 14. Alternative nucleic acid sequences for loading are described in U.S. Patent Publication No. 2010 / 0297699. and the entire contents of which are incorporated herein. Thus, in certain embodiments, cell thrombin The cells in the chamber device contain the nucleic acid sequence set forth in SEQ ID NO: 11 and / or SEQ ID NO: In some embodiments, the cell chamber device may comprise a nucleic acid sequence as described in 12. The cells in the device contain nucleic acids comprising the nucleic acid sequence of SEQ ID NO: 13 and the nucleic acid sequence of SEQ ID NO: 14. .

[0143] In some embodiments, the cells in the cell chamber device disclosed herein are α4β 7. Express one or more nucleic acids encoding an antibody that specifically binds to 7, or an antigen-binding portion thereof. In certain embodiments, the cells are vedolizumab or vedolizumab antigen-binding In certain embodiments, the cell chamber deactivates one or more nucleic acids encoding the portion. The cells in the device contain the nucleic acid sequence shown in SEQ ID NO: 11 and / or the nucleic acid sequence shown in SEQ ID NO: 12. In some embodiments, the cells in the cell chamber device express a nucleic acid sequence comprising sequence no. The nucleic acid comprising the nucleic acid sequence of SEQ ID NO: 13 and the nucleic acid sequence of SEQ ID NO: 14 is expressed.

[0144] In some embodiments, the cells in the cell chamber device contain a signal peptide. The nucleic acid sequence of the immature humanized anti-α4β7 immunoglobulin chain (e.g., SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, or SEQ ID NO: 20) In other embodiments, the cells in the cell chamber device express a signal peptide. expressing a nucleic acid comprising the nucleic acid sequence of a mature humanized anti-α4β7 immunoglobulin chain (e.g., For example, nucleotides 77 to 1429 of SEQ ID NO: 15 and nucleotides 79 to 7 of SEQ ID NO: 16 35, nucleotides 76 to 1428 of SEQ ID NO: 17, nucleotides 78 to 1428 of SEQ ID NO: 18 734, nucleotides 58 to 714 of SEQ ID NO: 19, or nucleotides of SEQ ID NO: 20 58~1410).

[0145] In one embodiment, the cells in the cell chamber device contain a recombinant expression vector and The expression vector comprises a first nucleic acid encoding an immunoglobulin heavy chain and a second nucleic acid encoding an immunoglobulin light chain. and a second nucleic acid encoding 1429, and the second nucleic acid comprises nucleotides 79 to 735 of SEQ ID NO:16.

[0146] In one embodiment, the cells in the cell chamber device contain a recombinant expression vector and The expression vector comprises a first nucleic acid encoding an immunoglobulin heavy chain and a second nucleic acid encoding an immunoglobulin light chain. and a second nucleic acid encoding 1428, and the second nucleic acid comprises nucleotides 78 to 734 of SEQ ID NO:18.

[0147] In one embodiment, the cells in the cell chamber device contain a recombinant expression vector and The expression vector comprises a first nucleic acid encoding an immunoglobulin heavy chain and a second nucleic acid encoding an immunoglobulin light chain. and a second nucleic acid encoding 1410, and the second nucleic acid comprises nucleotides 58 to 714 of SEQ ID NO:19.

[0148] Additional examples of nucleic acid sequences that can be expressed by cells in a cell chamber device are also described in WO 2008 / 115504, the entire contents of which are incorporated herein by reference. do.

[0149] In an exemplary embodiment, the cell chamber device comprises a polymerase comprising the amino acid sequence of SEQ ID NO:1. an antibody comprising a light chain variable region and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 5, or In some embodiments, the cell channel comprises an ARPE-19 cell that secretes an antigen-binding portion of the antibody. The device comprises CDR1 of SEQ ID NO: 2, CDR2 of SEQ ID NO: 3, and CDR3 of SEQ ID NO: 4. 3, and a heavy chain variable region comprising CDR1 of SEQ ID NO: 6, CDR2 of SEQ ID NO: 7, and A human antibody or antigen-binding portion thereof, which comprises a light chain variable region containing CDR3 of sequence number 8. In some embodiments, the cell chamber device comprises ARPE-19 cells comprising SEQ ID NO: an antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 9 and a light chain comprising the amino acid sequence of SEQ ID NO: 10. In some embodiments, the antibody comprises an ARPE-19 cell that secretes the antibody, or an antigen-binding portion thereof. The cell chamber device comprises a nucleic acid sequence comprising the nucleic acid sequence of SEQ ID NO:11 and the nucleic acid sequence of SEQ ID NO:12. In some embodiments, the cell chamber device comprises ARPE-19 cells containing an acid. ARPE-19 cells containing a nucleic acid comprising the nucleic acid sequence of SEQ ID NO: 13 and the nucleic acid sequence of SEQ ID NO: 14 In certain embodiments, the cell chamber device comprises an A that secretes vedolizumab. Contains RPE-19 cells.

[0150] In some embodiments, the cells in the cell chamber devices disclosed herein are expressing one or more nucleic acids encoding an antibody, or an antigen-binding portion thereof, that specifically binds to α In certain embodiments, the cells in the cell chamber device are selected from the group consisting of the cells shown in SEQ ID NO: 42. and / or the nucleic acid sequence shown in SEQ ID NO: 43. wherein the cells in the cell chamber device are harboring the nucleic acid sequence of SEQ ID NO: 44 and / or SEQ ID NO: In one embodiment, the cells in the cell chamber device express a nucleic acid comprising the nucleic acid sequence of 45. The recombinant expression vector comprises a vector encoding an immunoglobulin heavy chain. and a second nucleic acid encoding an immunoglobulin light chain, wherein the first nucleic acid , SEQ ID NO:44, and the second nucleic acid comprises SEQ ID NO:45.

[0151] In some embodiments, the cells in the cell chamber devices disclosed herein are IL- 12, or one or more nucleic acids encoding an antibody or antigen-binding portion thereof. In certain embodiments, the cells in the cell chamber device express the nucleotide sequence set forth in SEQ ID NO:46. and / or the nucleic acid sequence shown in SEQ ID NO: 47. In this embodiment, the cells in the cell chamber device contain the nucleic acid sequence of SEQ ID NO: 48 and / or SEQ ID NO: In one embodiment, the cells in the cell chamber device express a nucleic acid comprising the nucleic acid sequence of SEQ ID NO:49. The cell contains a recombinant expression vector, the recombinant expression vector encoding an immunoglobulin heavy chain. and a second nucleic acid encoding an immunoglobulin light chain, The first nucleic acid comprises SEQ ID NO:48 and the second nucleic acid comprises SEQ ID NO:49.

[0152] In certain embodiments, the cells in the cell chamber device are used to treat short bowel syndrome. The peptides can secrete peptide therapeutics for gastrointestinal use, such as: Examples of peptide therapeutics useful for this purpose are those described in US Pat. No. 9,125,882 (e.g., SEQ ID NO: 54). , US 9,742,455 (e.g., SEQ ID NO: 1), and US 7,737,251 (e.g., For example, SEQ ID NO: 8. Each of the foregoing patents and patent applications is incorporated herein by reference. It is incorporated herein in its entirety.

[0153] The amino acid sequence of an exemplary peptide therapeutic for gastrointestinal use is set forth as SEQ ID NO:21. In certain embodiments, the cells in the cell chamber are selected from the group consisting of SEQ ID NO:2. In some embodiments, the polypeptides disclosed herein secrete a peptide having an amino acid sequence of The cells in the cell chamber device express a peptide having the amino acid sequence of SEQ ID NO: 21. It includes nucleic acid encoding it.

[0154] In certain embodiments, the cell chamber device comprises the amino acid sequence of SEQ ID NO: 21. In some embodiments, the cell chamber comprises ARPE-19 cells secreting a peptide that binds to the cell. The device comprises an AR gene encoding a peptide having the amino acid sequence of SEQ ID NO:21. Contains PE-19 cells.

[0155] The cells are transfected by inserting into the cells an expression construct encoding the biomolecule using standard techniques. Thus, they can be engineered to produce biomolecules such as those described herein. For example, a vector containing a polynucleotide encoding a polypeptide of interest is inserted into a cell. By using the vector, a cell line that produces the desired polypeptide can be produced. The term, as used herein, refers to a gene that is capable of transferring genetic material into a cell and after which it can be replicated and / or integrated into the cellular genome and expressed, e.g. , is intended to refer to a nucleic acid molecule. One type of vector is a plasmid, It refers to a circular double-stranded DNA to which additional DNA segments can be ligated. Common vectors include phage vectors and viral vectors. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., , episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors). may be integrated into the genome of the host cell upon introduction into the host cell, thereby Moreover, certain vectors can replicate together with genes to which they are operatively linked. Such vectors are referred to herein as "recombinant expression vectors." ", or simply "expression vector" or "expression construct." Therapeutic Protein A variety of vectors suitable for recombinant expression of genes, e.g., therapeutic antibodies, are publicly available. Vector components generally include a signal sequence, an origin of replication, one or more one or more of a marker gene, an enhancer element, a promoter, and a transcription termination sequence of the Any signal sequence, origin of replication, etc. that may be used. Marker genes, enhancer elements, and transcription terminator sequences are well known in the art. and is described in further detail, for example, in U.S. Patent No. 7,053,202. In certain cases, the promoter element that drives the expression of the biomolecule is Constitutive promoter elements to ensure long-term expression of biomolecules. The present invention also provides a method for the production of a biomolecule by, for example, increasing the copy number of a nucleic acid that expresses the biomolecule. by removing premature termination or splicing signals within the nucleic acid that expresses the gene. Select a promoter for the site of genomic integration that allows for increased expression of the biomolecule, or and by various other methods known in the art. Another example of increasing expression levels is by empirically determining, for example, by genome sequencing. or transcriptional hopping by homologous recombination, computationally determined by expression probability algorithms. The target is incorporated into the target spot.

[0156] IV. Three-dimensional cell culture Also provided herein are nanofibers capable of accommodating three-dimensional cell cultures. and a cell chamber device comprising a polymer (e.g., electrospun). Thus, in some embodiments, the device may comprise a tissue or portion thereof having a three-dimensional structure (e.g., tissue explants), or clusters of cells (e.g., organoids or spheroids) A cell chamber device suitable for three-dimensional cell culture may include, for example, In methods of treatment where secretion of a biomolecule endogenously produced by a cell or tissue is desired, Cells or tissues may be cultured in accordance with methods known in the art. and grown in 3D cell culture (i.e., 3D cell culture) (e.g., as described herein by reference). Assay and drug development technologies,12(4),207 3D structures (see, e.g., ref. 218) can then be added to the cell chamber device. Cells or tissues having the phenotype can be isolated using art-recognized techniques, e.g., by isolating the tissue. Obtained from explants, biopsies, or harvested from living donors, cell cultures, or autopsies It is possible.

[0157] In some embodiments, the cell chamber device is a three-dimensional (3D) cell chamber that contains a tissue, or a portion thereof. Examples of tissues that can be included in the devices herein include liver tissue, pancreatic tissue, and the like. The tissue may optionally be a tissue explant (e.g., a tissue from a mouse, a mammal, a mammalian cell ... In one embodiment, the tissue or organ may be a piece or pieces of tissue or organ removed from an animal. In one embodiment, the cell chamber contains liver tissue (e.g., liver tissue explants). The alveolar chamber contains pancreatic tissue (e.g., pancreatic tissue explants such as islet tissue explants). In one embodiment, the cell chamber comprises kidney tissue (e.g., a kidney tissue explant). In some embodiments, the cell chamber comprises reproductive tissue (eg, ovarian or testicular tissue). In some embodiments, the tissue is a human kidney tissue explant, a human liver tissue explant, or a human pancreatic tissue explant. In other embodiments, the tissue is a human tissue, such as a human ovarian or human testicular explant. and other human reproductive tissues.

[0158] Alternatively, the cell chamber device may contain clusters of cells having a three-dimensional structure. For example, in some embodiments, the three-dimensional cell culture is an organoid or spheroid. In some embodiments, clusters of cells (e.g., organoids) can be included. or spheroids) can be cultured with hepatocytes (e.g., human hepatocytes), kidney cells, or Langerhans cells. Clusters of cells (e.g., organoids or spheroids) contain islet cells. To maintain the three-dimensional structure of the cell culture, the cells may be organized around a structure. In embodiments, the cell chamber comprises a set of cells organized around a structure such as a tube or sinusoid. Includes woven groups.

[0159] A cell chamber device for use with three-dimensional cell culture is described herein. As described above, the present invention provides a multilayer scaffold comprising a nanofibrous, e.g., electrospun, polymer. (See, e.g., Section II). Optionally, the scaffold layer is The medicament may be tailored to promote angiogenesis towards tissues or cells of interest.

[0160] In an alternative embodiment, the cell chamber device comprises a single nanofibrous, e.g., electro The single layer scaffold may comprise a polymer layer of Trospan, polyester, polyethylene Polybutylene Terephthalate (PET, also known as Dacron) Various polymers, such as polybutadiene (PBT), or polyurethane (PU), e.g., nanofibrous For example, in some embodiments, the monolayer scaffold may be formed from a nano- Fibrous polyester, nanofibrous polyethylene terephthalate (nPET), nanofibrous Polybutylene terephthalate (nPBT), and / or nanofibrous polyurethane (nP In one embodiment, the monolayer scaffold comprises nPET. In one embodiment, the monolayer scaffold comprises nPBT. In one embodiment, the monolayer scaffold comprises nPET-nPBT. In a further embodiment, the monolayer scaffold comprises nPU. , using a polymer sheet comprising a single layer, e.g., a homogeneous layer, of nanofibrous polymer, Generally, the multilayer devices can be fabricated as described with respect to the multilayer devices provided herein. In some embodiments, a single layer device may have a reduced pressure within the chamber compared to a multi-layer scaffold device. This can allow for greater contact between the cells and the environment outside the chamber.

[0161] In some embodiments, the nanofibrous polymer scaffold can be loaded with an anti-inflammatory agent. For example, the nanofibrous polymer scaffold may be loaded with tacrolimus, pirfenidone, and / or rofluridone. Without wishing to be bound by theory, it is believed that the anti-inflammatory The inclusion of an agent inhibits the infiltration of cells (e.g., immune or inflammatory cells) from outside the chamber. Inhibiting the growth of cells may help preserve the three-dimensional structure of cells growing within the chamber. The inhibitor may also reduce the subject's immune response to the device after implantation. In some embodiments, the cell chamber device comprises a monolayer scaffold comprising a nanofibrous polymer. The scaffold may be coated with anti-inflammatory drugs (e.g., tacrolimus, pirfenidone, and / or roflumin In another embodiment, the cell chamber device comprises a nanofibrous polymer. and a multi-layer scaffold comprising an anti-inflammatory agent (e.g., tacrolimus, pirfenidone, and / or a steroid). or roflumilast).

[0162] The tissue graft or cells having a three-dimensional structure can optionally secrete biomolecules. In some embodiments, the biomolecule is a biomolecule that is endogenously produced by a tissue or cell. In an alternative embodiment, the cells loaded into the cell chamber device of the present disclosure genetically engineered cells that have been engineered to produce, e.g., secrete, a biomolecule of interest; For example, they may be recombinant cells. Biomolecules that may be secreted by cells of the device include: For example, but not limited to, polypeptides, polysaccharides, and polynucleotides, and lipids (e.g., phospholipids, glycolipids, and sterols), chemical messengers (e.g., , neurotransmitters, and hormones such as insulin), vitamins, sugars (e.g., carbohydrates, Disaccharides, oligosaccharides, polysaccharides), amino acids, peptides, oligopeptides, polypeptides, proteins Proteins, nucleotides, deoxyribonucleic acid (DNA), or ribonucleic acid (RNA) Other secreted biomolecules include exosomes, lipid polymers, and These may include those assembled, packaged, and secreted as viral particles. In embodiments, the tissue graft or cells are administered with a therapeutic biomolecule as described herein. To secrete.

[0163] In some embodiments, tissues or cells (e.g., cells having a three-dimensional structure) within the device. ) can be used to synthesize one or more proteins or peptides, e.g., one or more therapeutic proteins or For example, tissue or cells within the device secrete an antibody or its antigen. Binding fragments, growth factors, hormones (e.g., insulin), cytokines, clotting factors (e.g., For example, factor VIII or factor IX, or variants thereof, e.g., Recombinase inate, Kogenate, Refacto, Advate, Alprolix,B eneFIX, Rixubis, Ixinity, Idelvion, etc.), or The polypeptides may secrete one or more therapeutic proteins, such as a combination of these.

[0164] Proteins secreted by tissues or cells within the device may, in some cases, For example, in one embodiment, the protein may be a protein that is endogenously produced by a tissue or cell. In the embodiment, the cell chamber device contains pancreatic tissue or pancreatic cells having a three-dimensional structure (e.g., Pancreatic tissue or cells contain insulin, amylin, glucagon, Secrete somatostatin, ghrelin, and / or other metabolic enzymes. In this embodiment, the cell chamber contains pancreatic tissue or cells (e.g., Langerhans cells) having a three-dimensional structure. Pancreatic tissue or cells include pancreatic islet cells, which secrete insulin. In this embodiment, the cell chamber is made of reproductive tissue (e.g., ovarian tissue or testicular tissue), or The reproductive tissue or cells derived therefrom may be from a subject to which the device is administered. secrete hormones or agents with hormone-regulating properties.

[0165] In other embodiments, the cell chamber device contains cells that perform functions such as detoxification or metabolism. It may contain cells.

[0166] In another embodiment, the cell chamber device is a three-dimensional structure of liver tissue or liver cells. Liver tissue or hepatocytes contain albumin, transferrin, plasminogen secrete fibrinogen, α-fetoprotein, and / or clotting factors. In embodiments, the liver tissue or liver cells are enriched in bile, alanine transaminase (ALT), Aspartate transaminase (AST), alkaline phosphatase (ALP), and Digestive enzymes such as gamma-glutamyl transpeptidase (GGT) and / or It can secrete substances.

[0167] In another embodiment, the cell chamber device comprises a renal tissue or renal cells having a three-dimensional structure. The renal tissue or cells contain erythropoietin, calcitriol, prostaglandins, secrete vasodilators, vasodilators, and / or renin.

[0168] In some embodiments, the tissue or cells in the device contain one protein or peptide. A gene capable of secreting, for example, one endogenous or recombinant protein or peptide. In other embodiments, the cells in the device express more than one protein, e.g., 2, 3, capable of secreting 4, 5, 6, 7, 8, 9, 10 or more endogenous or recombinant proteins In some embodiments, the device secretes multiple proteins or peptides. , a single tissue or cell cluster having a three-dimensional structure, e.g., a single tissue or cell Clusters can be loaded.

[0169] V. Methods of Administration and Treatment Also provided herein is a method for delivering a biomolecule to a subject, comprising administering to the subject: 1. A method comprising administering a cell chamber device as disclosed herein, wherein the device The cell chamber contains cells that secrete biomolecules, such as those described above. In certain cases, the recombinant peptide or protein may comprise The method can be used to detect antibodies, such as, for example, anti-α4β7 antibodies (e.g., vedolizumab), or The antigen-binding portion can be delivered to a subject. The dose of biomolecules delivered by the device depends on the dimensions of the cell chamber device (length, diameter, volume), and modulating the number of cells within the device through geometry; Regulating the expression level of a biomolecule by a cell (e.g., altering copy number, promoter by selecting a device), and / or adjusting the number of devices delivered to the subject ( For example, 1 to 10 devices per patient can be controlled. The elephants may be administered one or more of the cell chamber devices described herein. In some embodiments, the subject is administered one cell chamber device. In this case, the subject is asked to throw 2, 3, 4, 5, 6, 7, 8, 9, 10 or more cell chamber devices. Multiple devices can be administered at the same (or nearly the same) site or or at multiple sites within the body.

[0170] Prior to administration of the device, the device is loaded with cells that secrete the biomolecules to be delivered to the subject. The cells are loaded, for example, through an opening or loading port in the scaffold surrounding the chamber. The device can be loaded by injecting cells into the cell chamber. Alternatively, the device can be pre-loaded with cells. The number of cells in the device administered to a subject may vary depending on the dimensions of the device, The amount of biomolecule secreted by the cells and the desirability of the biomolecule to be delivered to the recipient subject. The amount of biomolecules secreted from the device per day may vary depending on the dosage. mg or the desired concentration of the biomolecule in the serum or plasma of the recipient subject The desired number of cells in the cell chamber device can be determined by methods known in the art. and according to the methods outlined herein (see, e.g., Example 1). For example, the number of biomolecules secreted per cell per day can be easily determined. Quantification can be performed using standard cell count and biomolecule quantification assays (e.g., Western blot or E Based on this determination, the size of the cell chamber can be measured. The size of the cells in the cell chamber device (i.e., the number of cells in the device after growth and saturation) can be adjusted. the number of cells in the device) and therefore the dose of biomolecules secreted by the device per day. The cells in the chamber can grow until they reach the capacity of the device. Therefore, the number of cells seeded onto the device can be adjusted to the number of cells in the device after implantation. It may be less than that.

[0171] In some embodiments, the device is administered in a concentration of about 1×10 4 cells ~ approx. 1 x 10 1 2 cells (e.g., approximately 1 x 10 4 cells ~ approx. 1 x 10 5 cells, approximately 1 x 10 5 pieces cells ~ approx. 1 x 10 6cells, approximately 1 x 10 6 cells ~ approx. 1 x 10 7 cells, approximately 1 x 1 0 7 cells ~ approx. 1 x 10 9 cells, or approximately 1 x 10 9 cells ~ approx. 1 x 10 12 pieces In some embodiments, the device is loaded with, and therefore can contain, cells. The chair is approximately 1 x 10 6 cells ~ approx. 1 x 10 7 cells (e.g., approximately 1 x 10 6 individual cells ~Approx. 1×10 7 cells, approximately 2 x 10 6 cells ~ approx. 1 x 10 7 cells, approximately 3 x 10 6 cells ~ approx. 1 x 10 7 cells, approximately 4 x 10 6 cells ~ approx. 1 x 10 7 cells, approximately 5 x10 6 cells ~ approx. 1 x 10 7 cells, approximately 6 x 10 6 cells ~ approx. 9 x 10 6 Individual details Cells, approximately 7 x 10 6 cells ~ approx. 9 x 10 6 cells, or approximately 8 x 10 6 cells ~ approximately 9 x10 6 For example, in certain embodiments, the device comprises about 8.5× 10 6 In some embodiments, the device contains about 1 x 10 cells. 7 cells ~ approx. 1 x10 8 In some embodiments, the device contains about 1 x 10 cells. 8 ~approximately 100 cells 1×10 9 In some embodiments, the device contains about 1 x 10 cells. 9 Individual cells Approximately 1×1010 In some embodiments, the device contains about 1 x 10 cells. 10 pieces cells ~ approx. 1 x 10 11 In some embodiments, the device contains about 1 x 10 cells. 1 1 cells ~ approx. 1 x 10 12 In an exemplary embodiment, the device contains about 1 x10 4 cells, 1 x 10 5 cells, 1 x 10 6 cells, 1 x 10 7 cells, 1 x10 8 cells, 1 x 10 9 cells, 1 x 10 10 cells, 1 x 10 11 individual cells , or 1 × 10 12 In some embodiments, the device comprises a cell. Before filling, about 1 x 10 4 cells, 1 x 10 5 cells, 1 x 10 6 cells, 1×1 0 7 cells, 1 x 10 8 cells, 1 x 10 9 cells, 1 x 10 10 cells, 1x 10 11 cells, or 1 x 10 12 Load cells.

[0172] In some embodiments, the device has a capacitance of about 1×10 4 cells / cm 2 ~Approx. 1×10 6 pieces cells / cm 2 , 1×10 5 cells / cm 2 ~Approx. 1×10 6 cells / cm 2 , about 1×1 0 5 cells / cm 2~Approx. 9×10 5 cells / cm 2 , about 2×10 5 cells / cm 2 ~Approx. 8×10 5 cells / cm 2 , about 3×10 5 cells / cm 2 ~Approx. 7×10 5 Individual details cells / cm 2 , about 5×10 5 cells / cm 2 ~Approx. 7×10 5 cells / cm 2 , about 6×1 0 5 cells / cm 2 ~Approx. 7×10 5 cells / cm 2 , about 6×10 5 cells / cm 2 ~Approx. 6.5×10 5 cells / cm 2 , or 6.25 × 10 5 cells / cm 2 Contains .

[0173] In other embodiments, the device may be used after implantation in a subject (e.g., after the cells in the chamber have grown). After a period sufficient for reproduction, approximately 1 × 10 4 cells ~ approx. 1 x 10 12 Individual cells (e.g. If so, it is about 1 × 10 4 cells ~ approx. 1 x 10 5 cells, approximately 1 x 10 5 cells ~ approx. 1 x 10 6 cells, approximately 1 x 10 6 cells ~ approx. 1 x 10 7 cells, approximately 1 x 10 7 ~approximately 100 cells 1×10 9 cells, or approximately 1 x 10 9 cells ~ approx. 1 x 10 12 Contains 100 cells In some embodiments, the device has a capacity of about 1×10 6 cells ~ approx. 1 x 10 7 cells (e.g., approximately 1 x 10 6 cells ~ approx. 1 x 10 7 cells, approximately 2 x 10 6 pieces cells ~ approx. 1 x 10 7 cells, approximately 3 x 10 6 cells ~ approx. 1 x 10 7 cells, approximately 4 x 1 0 6 cells ~ approx. 1 x 10 7 cells, approximately 5 x 10 6 cells ~ approx. 1 x 10 7 individual cells, Approximately 6×10 6 cells ~ approx. 9 x 10 6 cells, approximately 7 x 10 6 cells ~ approx. 9 x 10 6 pieces cells, or approximately 8 x 10 6 cells ~ approx. 9 x 10 6 can accommodate 100 cells For example, in one particular embodiment, the device has a capacity of about 8.5×10 6 Contains cells In some embodiments, the device can have a capacity of about 1×10 7 cells ~ approx. 1 x 10 8 In some embodiments, the device can accommodate approximately 1 x 10 cells. 8 pieces cells ~ approx. 1 x 10 9 In some embodiments, the device can accommodate cells. , about 1×10 9 cells ~ approx. 1 x 10 10 Some implementations can accommodate cells. In morphology, the device has a capacitance of approximately 1 x 10 10 cells ~ approx. 1 x 10 11 It can accommodate 100 cells. In some embodiments, the device can have a capacity of about 1×10 11cells ~ approx. 1 x 10 1 2 In an exemplary embodiment, the device can accommodate approximately 1 x 10 cells. 4 cells, 1 x 10 5 cells, 1 x 10 6 cells, 1 x 10 7 cells, 1 x 10 8 cells, 1 x 10 9 cells, 1 x 10 10 cells, 1 x 10 11 cells, or 1×10 12 In some embodiments, the device can accommodate a number of cells of interest. After embedding in , approximately 1 × 10 4 cells, 1 x 10 5 cells, 1 x 10 6 individual cells, 1×10 7 cells, 1 x 10 8 cells, 1 x 10 9 cells, 1 x 10 10 individual cells , 1×10 11 cells, or 1 x 10 12 Load cells.

[0174] The implanted device may be inserted into the subject, for example, through a surgical incision at the site of implantation. The site of implantation will depend on the configuration of the device, the disorder to be treated, and This can vary depending on the desired biodistribution of biomolecules secreted by cells within the device. In some embodiments, the device is placed under the skin (subcutaneous implantation), on the omentum, or in or on the liver. The subject is administered by implantation at a site selected from a site adjacent to the liver. In embodiments, the cell chamber device is located on or under the skin; on a mucosal surface, in a body cavity, in the peritoneal cavity; the central nervous system, e.g., the brain, cortex, ventricles, or spinal cord; organs, e.g., the heart, liver, kidneys; Spleen, lungs, pancreas, lymphatic system, vascular system, oral cavity, nasal cavity, teeth, gums, GI tract; bones; hip joints p); adipose tissue; muscle tissue; circulating blood; eyes (e.g., intraocular); breasts, vagina; uterus, joints, e.g. For example, they are implanted in the knee or hip joint, or in the spine. In embodiments, the cell chamber device may be used to detect specific parts or tissues of the body, e.g., blood, Eyes, brain, skin, lungs, stomach, mouth, ears, legs, feet, hands, liver, heart, kidneys, bones, reproductive organs, testes, In some embodiments, the cells are implanted into the pancreas, spleen, large intestine, small intestine, spinal cord, or muscle. The chamber device may be used to control certain systems of the body, such as the vascular system, the nervous system (e.g., peripheral nervous system (PNS) or central nervous system (CNS), skeletal system, respiratory system, endocrine system, lymphatic In some embodiments, the biomolecule is administered to deliver the biomolecule to the uterine, reproductive, or gastrointestinal tract. The cell chamber device may be placed under the subject's skin, on the subject's omentum, in the subject's subcutaneous fat, or The device is implanted in or adjacent to the muscle tissue of a subject. The device can be implanted subcutaneously.

[0175] In some embodiments, the cell chamber is implanted in the peritoneal cavity (e.g., omentum). In an embodiment, the cell chamber is a bursa lesser omentalis (also known as an omental bursa or (also known as bursalis omentum), greater omentum, lesser omentum, stomach implanted in or on the small intestine, large intestine, liver, spleen, gastrosplenic ligament, adrenal gland, or pancreas do.

[0176] The cell chamber device may, in some cases, cause injury to the subject, e.g. be easily retrieved from the subject without causing significant damage to the surrounding tissue In one embodiment, the device can be implanted in a minimally invasive manner, e.g. Minimal dislodgement of the device from surrounding tissue via surgical approach, extraction, or excision It can be retrieved without disciplinary or surgical separation.

[0177] The length of time the implant remains in the subject will depend on the condition being treated and the nature of the device. The therapeutic dose of the biomolecules secreted by the cells will vary depending on the In some embodiments, the cell chamber device can be used to culture the cells in a subject for at least 2, 3, 4, 5 days. , 6 days, 7 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 10 weeks, 12 weeks, 1 month , 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months months, 20 months, 21 months, 22 months, 23 months, 24 months, 1 year, 1.5 years, 2 years, 2 .5 years, 3 years, 3.5 years, or 4 years, 4.5 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years In some embodiments, the cells are maintained for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or more years. The cell chamber device was implanted in the subject for approximately 1-2 days, 1-7 days, 2- 4 days, 3-5 days, 4 days-1 week, 1-2 weeks, 1-4 weeks, 2-4 weeks, 2 weeks to 6 weeks, 3 weeks to 5 weeks, 4 weeks to 1 month, 1 month to 2 months, 1 month to 4 months, 1 month to 6 months, 2 months to 4 months, 3 months to 5 months, 4 months to 6 months, 5 months to 7 months, 6-8 months, 6-12 months, 7-9 months, 8-10 months, 9-11 months months, 10-12 months, 11-13 months, 12-15 months, 15-18 months months, 18-21 months, 21-24 months, 2-2.5 years, 2.5-3 years, 3 Years ~ 3.5 years, 3.5 years ~ 4 years, 4 years ~ 5 years, 5 years ~ 7 years, 7 years ~ 9 years, 9 years ~ 11 years, It can be maintained for an average period of 11 to 13 years, or 13 to 15 years. In some embodiments, the device remains in the subject for at least 30 days. In certain embodiments, the device is implanted in the subject for at least 90 days. In certain embodiments, the device is implanted in the subject for at least 120 days. The device is implanted in the subject for at least one year. In certain embodiments, the device and implanted in the subject for at least two years. In certain embodiments, the device In certain embodiments, the device is implanted in the subject for at least three years. In certain embodiments, the device is implanted in the subject for at least 7 years. In certain embodiments, the device is implanted in the subject for at least 10 years. In certain embodiments, the device is implanted in the subject for at least 12 years. In certain embodiments, the device is implanted in the subject for at least 15 years. In an embodiment, the device is permanently implanted in the subject.

[0178] In some embodiments, the cell chamber device achieves a therapeutic benefit in a subject. for a period of time effective to (e.g., achieve a therapeutically effective plasma concentration of the biomolecule in the subject); of biomolecules (e.g., recombinant peptides or proteins) secreted by cells The daily dose is at least 1 mg / day (e.g., at least about 1 mg / day, at least about 2 mg / day, at least about 3 mg / day, at least about 4 mg / day, at least about 4.5 mg / day, at least about 5 mg / day, at least about 6 mg / day, at least about 7 mg / day, at least about 8 mg / day, at least about 9 mg / day, or greater than 9 mg / day) In some embodiments, the cell chamber device is maintained in the subject. achieve a therapeutically effective plasma concentration of a biomolecule in a subject ) for a period of time effective to produce a biomolecule (e.g., a recombinant peptide or a recombinant protein) secreted by the cell. The daily dose of recombinant protein is 0.5mg / day to 1mg / day, 1mg / day to 2mg / day , 1mg / day~5mg / day, 2mg / day~5mg / day, 4mg / day~7mg / day, 5mg / day~8mg / day, 5mg / day~10mg / day, 6mg / day~9mg / day, 7mg / day~ 10mg / day, 10mg / day to 15mg / day, 8mg / day to 11mg / day, 6mg / day and above The dose is maintained within the subject at 12 mg / day, or between 9 mg / day and 12 mg / day.

[0179] For example, in some embodiments, the cell chamber device may be configured to contain a biomolecule (e.g., a recombinant peptide). The plasma concentration of the peptide or recombinant protein (or peptides) is increased in a subject over a period of time after implantation, e.g. For example, for a period of time effective to achieve a therapeutic benefit in a subject, e.g., at least 2 weeks after implantation. At least 5 days, at least 10 days, at least 20 days, at least 30 days, at least 40 days, at least 50 days, at least 60 days, at least 3 months, at least 4 months, at least At least 5 months, at least 6 months, at least 12 months, 1-2 days, 1-7 days, 2 days ~4 days, 3-5 days, 4 days to 1 week, 1 week to 2 weeks, 1 week to 4 weeks, 2 weeks to 4 weeks , 2 weeks to 6 weeks, 3 weeks to 5 weeks, 4 weeks to 1 month, 1 month to 2 months, 1 month to 4 months , 1 month to 6 months, 2 months to 4 months, 3 months to 5 months, 4 months to 6 months, 5 months to 7 months , 6 months to 8 months, 6 months to 12 months, 7 months to 9 months, 8 months to 10 months, 9 months to 1 1 month, 10-12 months, 11-13 months, 12-15 months, 15 months to 1 8 months, 18-21 months, 21-24 months, 2-2.5 years, 2.5-3 years, 3 to 3.5 years, or 3.5 to 4 years, at least 5 μg / mL (e.g., at least about 5 μg / mL, at least about 10 μg / mL, at least about 15 μg / mL, at least at least about 17 μg / mL, at least about 20 μg / mL, or greater than 20 μg / mL Thus, it is maintained within the subject.

[0180] In some embodiments, the cell chamber device may contain a biomolecule (e.g., a recombinant peptide or The plasma concentration of the target protein (or recombinant protein) is increased in a subject over a period of time after implantation, e.g., A period effective to achieve therapeutic benefit in the elephant, e.g., 1-2 days after implantation; 1-7 days, 2-4 days, 3-5 days, 4 days-1 week, 1-2 weeks, 1-4 weeks , 2 weeks to 4 weeks, 2 weeks to 6 weeks, 3 weeks to 5 weeks, 4 weeks to 1 month, 1 month to 2 months , 1 month to 4 months, 1 month to 6 months, 2 months to 4 months, 3 months to 5 months, 4 months to 6 months , 5 months to 7 months, 6 months to 8 months, 6 months to 12 months, 7 months to 9 months, 8 months to 10 months months, 9-11 months, 10-12 months, 11-13 months, 12-15 months months, 15-18 months, 18-21 months, 21-24 months, 2-2.5 years, 2.5 to 3 years, 3 to 3.5 years, or 3.5 to 4 years, 4 μg / mL to 8 μg / mL , 5μg / mL~9μg / mL, 5μg / mL~12μg / mL, 6μg / mL~10μ g / mL, 7μg / mL~11μg / mL, 7μg / mL~15μg / mL, 8μg / m L~12μg / mL, 9μg / mL~13μg / mL, 10μg / mL~15μg / mL , 15μg / mL to 20μg / mL, or 20μg / mL to 25μg / mL , and is maintained within the subject.

[0181] In some embodiments, the cell chamber device may contain a biomolecule (e.g., a recombinant peptide or or recombinant protein) in a subject after implantation, e.g., at least At least 5 days, at least 10 days, at least 20 days, at least 30 days, at least 40 days, At least 50 days, at least 60 days, at least 3 months, at least 4 months, at least At least 5 months, at least 6 months, at least 12 months, 1-2 days, 1-7 days, 2-4 days Days, 3-5 days, 4 days-1 week, 1-2 weeks, 1-4 weeks, 2-4 weeks, 2 1 week to 6 weeks, 3 weeks to 5 weeks, 4 weeks to 1 month, 1 month to 2 months, 1 month to 4 months, 1 months to 6 months, 2 months to 4 months, 3 months to 5 months, 4 months to 6 months, 5 months to 7 months, 6 6-8 months, 6-12 months, 7-9 months, 8-10 months, 9-11 months Months, 10-12 months, 11-13 months, 12-15 months, 15-18 months months, 18-21 months, 21-24 months, 2-2.5 years, 2.5-3 years, 3 years ~3.5 years, or 3.5 to 4 years, 1 μg / mL to 5 μg / mL, 1 μg / mL to 10 μg / mL, 5μg / mL~10μg / mL, 5μg / mL~15μg / mL, 10μg / mL~15μg / mL, 10μg / mL~20μg / mL, 15μg / mL~20μg / mL, 15μg / mL~30μg / mL, 20μg / mL~25μg / mL, 20μg / mL~40μg / mL, 25μg / mL~30μg / mL, 25μg / mL~50μg / mL, 30μg / mL~35μg / mL, 30μg / mL~60μg / mL, or 3 The concentration in the subject is maintained at 5 μg / mL to 40 μg / mL.

[0182] In some embodiments, the cell chamber device may contain a biomolecule (e.g., a recombinant peptide or The plasma concentration of the recombinant protein (or recombinant protein) in the subject is increased over a period of time after implantation, e.g., At least 5 days, at least 10 days, at least 20 days, at least 30 days, at least 4 0 days, at least 50 days, at least 60 days, at least 3 months, at least 4 months, At least 5 months, at least 6 months, at least 12 months, 1-2 days, 1-7 days, 2 7th-4th, 3rd-5th, 4th-7th, 7th-10th, 10th-20th, 20th-30th, 30-45 days, 45-60 days, 1-2 weeks, 1-4 weeks, 2-4 weeks, 2 weeks to 6 weeks, 3 weeks to 5 weeks, 4 weeks to 1 month, 1 month to 2 months, 1 month to 4 months, 1 month to 6 months, 2 months to 4 months, 3 months to 5 months, 4 months to 6 months, 5 months to 7 months, 6-8 months, 6-12 months, 7-9 months, 8-10 months, 9-11 months months, 10-12 months, 11-13 months, 12-15 months, 15-18 months months, 18-21 months, 21-24 months, 2-2.5 years, 2.5-3 years, 3 Between 3.5 years and 3.5 years, or between 3.5 years and 4 years, the mean serum creatinine concentration was at least 17 μg / mL. is maintained.

[0183] In some embodiments, the cell chamber device adjusts the dose of the biomolecule to 0.1 mg of the subject. / kg body weight to about 10.0 mg / kg body weight, for example, about 2 mg / kg to about 7 mg / kg, A period effective to deliver 3 mg / kg to about 6 mg / kg, or about 15 mg / kg to about 5 mg / kg In certain embodiments, the therapeutic agent is delivered over the course of the implant. The dose of the biomolecule is about 0.3 mg / kg, about 0.5 mg / kg, about 1 mg / kg, about 2 mg / kg, or mg / kg, approx. 3 mg / kg, approx. 4 mg / kg, approx. 5 mg / kg, approx. 6 mg / kg, approx. 7 mg / kg, about 8 mg / kg, about 9 mg / kg, or about 10 mg / kg.

[0184] In some embodiments, the implants are administered over a period of time, e.g., weekly, every two weeks, Total use of biomolecules delivered every 4, 6, 8, or 10 weeks The amounts are approximately 22 mg, approximately 50 mg, approximately 72 mg, approximately 100 mg, approximately 125 mg, and approximately 150 mg. g, approximately 165 mg, approximately 200 mg, approximately 300 mg, approximately 432 mg, approximately 450 mg, or In some embodiments, the implant may be administered at a dose of about 600 mg for a period of time, e.g. , per week, every 2 weeks, every 4 weeks, every 6 weeks, every 8 weeks, or every 10 weeks The total dose of biomolecule delivered is, for example, at least 77 mg, at least 125 mg In one embodiment, the implant may have a Period, e.g., weekly, every 2 weeks, every 4 weeks, every 6 weeks, every 8 weeks, or every 10 weeks In another embodiment, the total dose of biomolecule delivered over each period is 165 mg. The total dose every two weeks is 108 mg. In another embodiment, the total dose every two weeks is 21 In another embodiment, the total dose is 150 mg every two weeks. In one embodiment, the total dose is 200 mg every two weeks. In another embodiment, the total dose is 200 mg every four weeks. In another embodiment, the total dose is 300 mg every 8 weeks. For example, 300 mg of a biomolecule, e.g., anti-α4, per 8-week period, e.g., every 8 weeks. To deliver antibodies, such as β7 antibodies (e.g., vedolizumab), or antigen-binding portions thereof The implants are sized to deliver approximately 37.5 mg / week, which is In another example, the cells are loaded with cells capable of being injected per two week period, e.g., 1 every two weeks. 0.8 mg of a biomolecule, e.g., an antibody such as an anti-α4β7 antibody (e.g., vedolizumab), or or an antigen-binding portion thereof, the implant is designed to deliver approximately 54 mg / week. The foregoing examples are merely illustrative. and is not intended to be limiting.

[0185] In some embodiments, the dose of biomolecule delivered over the duration of the implant is 1 Approximately 0.5 mg / kg per week to 10 mg / kg per week, 2 mg / kg per week g to 6 mg / kg per week, 5 mg / kg per week to 15 mg / kg per week g, 10 mg / kg per week to 20 mg / kg per week, 15 mg per week / kg ~ 30mg / kg per week, 20mg / kg per week ~ 40mg / kg per week mg / kg per week, 30 mg / kg per week to 60 mg / kg per week.

[0186] The biomolecule is an antibody, e.g., an anti-α4β7 antibody (e.g., vedolizumab), or an antigen thereof. If the binding moiety is a The daily dose of the antibody, or antigen-binding portion thereof, administered is sufficient to achieve a therapeutic benefit in the subject. (e.g., achieve a therapeutically effective plasma concentration of the antibody, or antigen-binding portion thereof, in the subject) about 1 mg / day for a period effective to achieve a clinical benefit in a subject (e.g., at least about 1 mg / day, at least about 2 mg / day, at least about 3 mg / day) , at least about 4 mg / day, at least about 4.5 mg / day, at least about 5 mg / day, at least about 6 mg / day, at least about 7 mg / day, at least about 8 mg / day, at least The dose can be maintained in the subject at about 9 mg / day, or greater than 9 mg / day. In some embodiments, the cell chamber device comprises a chamber for detecting antibodies secreted by the cells, or The serum or plasma concentration of the antigen-binding moiety is effective to achieve a therapeutic benefit in the subject. for a suitable period of time, e.g., at least 30 days after implantation (e.g., at least 30 days, at least At least 45 days, at least 55 days, at least 60 days, at least 70 days, at least 90 days , at least 120 days, at least 150 days, at least 240 days, at least 365 days , or 30 to 90 days, 40 to 60 days, 50 to 70 days, 60 to 70 days, 60 days or more 90 days, 60-120 days, 65-75 days, 70-90 days, 90-120 days, 12 0-240 days, 240-365 days, or more than 365 days), at least about 5 μg / mL (e.g., at least about 5 μg / mL, at least about 10 μg / mL , at least about 15 μg / mL, at least about 17 μg / mL, at least about 20 μg / mL mL, or greater than 20 μg / mL).

[0187] In some embodiments, the cell chamber device comprises an antibody secreted by the cell, or for a period of time during which a daily dose of the antigen-binding portion is effective to achieve a therapeutic benefit in the subject; In subjects, 0.5 mg / day to 1 mg / day, 1 mg / day to 2 mg / day, 1 mg / day to 5 mg g / day, 2mg / day~5mg / day, 2mg / day~25mg / day, 4mg / day~7mg / day , 4mg / day~12mg / day, 5mg / day~8mg / day, 5mg / day~10mg / day, 6 mg / day~9mg / day, 7mg / day~10mg / day, 10mg / day~15mg / day, 8m g / day to 11 mg / day, 6 mg / day to 12 mg / day, or 9 mg / day to 12 mg / day As such, it is maintained within the subject.

[0188] In some embodiments, the cell chamber device comprises an antibody secreted by the cell, or The serum or plasma concentration of the antigen-binding portion is sufficient to achieve a therapeutic benefit in the subject. A validity period, e.g., at least 30 days after implantation (e.g., at least 30 days, At least 45 days, at least 55 days, at least 60 days, at least 70 days, at least 9 0 days, at least 120 days, at least 150 days, at least 240 days, at least 36 5 days, or 30-90 days, 40-60 days, 50-70 days, 60-70 days, 60 Days to 90 days, 60 days to 120 days, 65 days to 75 days, 70 days to 90 days, 90 days to 120 days, 120-240 days, 240-365 days, or over 365 days) 2.5μg / mL~7.5μg / mL, 4μg / mL~7μg / mL, 5μg / mL~8 μg / mL, 5μg / mL~10μg / mL, 5μg / mL~50μg / mL, 8μg / mL mL~15μg / mL, 10μg / mL~20μg / mL, 16μg / mL~20μg / mL, 18μg / mL~21μg / mL, 21μg / mL~30μg / mL, 25μg / mL mL to 35 μg / mL, or 31 μg / mL to 45 μg / mL be maintained.

[0189] In some embodiments, the cell chamber device contains a dose of an antibody, for example, anti-α4β7 The antibody (e.g., vedolizumab), or an antigen-binding portion thereof, is administered at about 0.1 mg / kg body weight to About 10.0 mg / kg body weight, for example, about 2 mg / kg to about 7 mg / kg, about 3 mg / kg ~ about 6 mg / kg, or about 15 to about 5 mg / kg, about 5 mg / kg body weight to about 25 mg / kg body weight, or about 10 mg / kg body weight to about 20 mg / kg body weight In certain embodiments, the therapeutic agent is delivered over the course of the implant. The total dose of the antibody (e.g., vedolizumab), or antigen-binding fragment thereof, is about 0.3 mg / kg, approx. 0.5mg / kg, approx. 1mg / kg, approx. 2mg / kg, approx. 3mg / kg, approx. 4m g / kg, approx. 5mg / kg, approx. 6mg / kg, approx. 7mg / kg, approx. 8mg / kg, approx. 9m g / kg, or about 10 mg / kg.

[0190] In some embodiments, the duration of the implant is, for example, about 2 weeks, about 4 weeks, about 6 weeks, Approximately 8 weeks, approximately 10 weeks, approximately 12 weeks, approximately 14 weeks, approximately 16 weeks, approximately 20 weeks, approximately 24 weeks , about 28 weeks, about 32 weeks, about 36 weeks or more of an antibody or antigen thereof The total dose of the binding fragment is about 22 mg, about 50 mg, about 72 mg, about 125 mg, about 165 mg. g, or approximately 432 mg, approximately 450 mg, approximately 300 mg, approximately 600 mg, approximately 650 mg, Approx. 900mg, Approx. 1200mg, Approx. 1300mg, Approx. 1500mg, Approx. 1800mg, Approx. In some embodiments, the dose may be 2100 mg or more over the duration of the implant. The total dose of antibody, or antigen-binding fragment thereof, delivered is at least 77 mg, at least In one embodiment, the implant may have a hydroxyl group of 125 mg, or at least 356 mg. The total dose of antibody, or antigen-binding fragment thereof, delivered over the period is 165 mg. In another embodiment, the total dose of the antibody, or antigen-binding fragment thereof, is 108 mg. In this embodiment, the total dose of the antibody, or antigen-binding fragment thereof, is 216 mg. In an embodiment, the total dose of the antibody, or antigen-binding fragment thereof, is 300 mg. Implants that stably deliver a total of three doses of 00 mg of biomolecule are typically 8 It is administered intravenously weekly, so it is implanted for 24 weeks and the device This will deliver a total of 900 mg (approximately 5.36 mg per day). In another example, an infusion was used to steadily deliver a total of six doses of 108 mg of a biomolecule. The plant is typically administered intravenously every two weeks, so that's 12 weeks. When implanted, the device will deliver a total of 650 mg (approximately 7.7 mg per day). mg).

[0191] In some embodiments, the antibody, or antigen thereof, delivered over the course of the implant. The dose of the binding moiety is about 0.5 mg / kg per week to 10 mg / kg per week, 2mg / kg per week to 6mg / kg per week, 5mg / kg per week to 1 week 15mg / kg per day, 10mg / kg per week to 20mg / kg per week, 15mg / kg per week to 30mg / kg per week, 20mg / kg per week 40 mg / kg per day, or 30 mg / kg per week to 60 mg / kg per week These dosages are exemplary and are not intended to be limiting. not present.

[0192] In a further aspect, provided herein is a method for administering to a subject a device as disclosed herein. a method for treating a subject having a disease by administering a cell therapy to a subject, the method comprising administering a cell therapy to a subject having a disease by ... The compound is administered in an amount and concentration suitable to treat the disease or ameliorate one or more symptoms of the disease. The cells secreting the desired biomolecule are those that are specific to the disease being treated. The treatment can be selected or manipulated based on the patient's condition.

[0193] Examples of diseases that can be treated according to the methods herein include gastrointestinal disorders, cancer, and respiratory disorders. , cardiovascular disease, neurological disease, autoimmune disorder, endocrine and / or metabolic disorder, blood disorder, if Commonly, eye disorders are included.

[0194] In some embodiments, the cell chamber device is used to treat inflammatory bowel disease or short bowel syndrome. Gastrointestinal disorders such as steroids, steroids, and steroid drugs can be treated according to the methods herein. Examples of inflammatory bowel diseases (IBD) include ulcerative colitis, Crohn's disease, and primary sclerosing biliary tract infection. inflammation, eosinophilic esophagitis, autoimmune hepatitis, ileitis, celiac disease, non-tropical sprue, serum Enteropathy associated with negative arthropathy, microscopic or collagenous colitis, eosinophilic gastroenteritis, or In some embodiments, inflammatory bowel disease may include pouchitis following rectal resection and ileoanal anastomosis. The intestinal disease is Crohn's disease or ulcerative colitis.

[0013] The present invention provides a method of treating a subject having a gastrointestinal disorder, comprising administering to the subject a compound disclosed herein. In certain such embodiments, the method comprises administering a device to treat gastrointestinal disorders. Subjects with gastrointestinal disorders are administered a device containing cells that secrete biomolecules that treat the gastrointestinal disorder. can be.

[0195] In some embodiments, the cell chamber device is adapted to treat a patient with inflammatory bowel disease or short bowel syndrome. In some embodiments, the cell channel is implanted into the peritoneum, mesenteric, or peritoneal space of a subject. The membrane device is inserted into the lesser omental bursa (i.e., the omentum) of a subject with inflammatory bowel disease or short bowel syndrome. (i.e., omental bursa or omental sac), in or on the greater omentum, lesser omentum, in or near the stomach, in the small intestine, The catheter is implanted in or near the large intestine, in or near the large intestine, or in the mesentery of the small intestine.

[0196] The cell chamber devices provided herein can be used to administer to a subject the cells contained therein. Any biomolecule secreted by a cell can be delivered. For example, as described above, Using the cell chamber device, protein or peptide therapeutics, as well as therapeutic antibodies The antibody, or an antigen-binding portion thereof, can be delivered. By way of example, the following embodiments are Exemplary treatments that can be achieved using the cell chamber devices described herein include: These exemplary embodiments illustrate methods for detecting and measuring the cell chamber temperature using the cell chamber device provided herein. The system can be used to deliver suitable biomolecules, e.g., therapeutic proteins, peptides, antibodies, etc. The present invention can be readily adapted to the treatment of other diseases or disorders through the These examples should not be construed as limiting in any way.

[0197] In one embodiment, provided herein is a method for treating a patient with Crohn's disease or ulcerative colitis. A method of treating a subject comprising administering to the subject a device disclosed herein. In certain such embodiments, Crohn's disease or ulcerative colitis is treated. Subjects with the disease may be treated with an antibody, e.g., an anti-α4β7 antibody (e.g., vedolizumab or abrilumab). In some embodiments, the patient is administered a device containing cells secreting the antigen-binding portion thereof. In one embodiment, the cell chamber device is configured to measure serum or plasma concentrations of antibodies, or antigen-binding portions thereof. The dose is effective to achieve a therapeutic benefit in the subject, e.g., for at least a few days after implantation. At least 30 days (e.g., at least 30 days, at least 45 days, at least 55 days, at least At least 60 days, at least 70 days, at least 90 days, 30 to 90 days, 40 to 60 days , 50-70 days, 60-70 days, 60-90 days, 60-120 days, 65-75 days days, 70-90 days, 90-120 days, 120-240 days, 240-365 days, at least 5 μg / mL (e.g., at least about 5 μg / mL, at least about 10 μg / mL, at least about 15 μg / mL, at least about 17 μg / mL, at least about 20 μg / mL, or greater than 20 μg / mL) In some embodiments, the medicament is maintained in a subject with Crohn's disease or ulcerative colitis. The cell chamber device is configured to deliver an antibody, or an antigen-binding portion thereof, to a subject. A period of time effective to achieve a therapeutic benefit, e.g., at least 60 days (e.g., 60 days) after implantation. 60-70 days, 65-75 days, 70-90 days, 90-120 days, 120 days or more 10 mg / week to 50 mg / week for a period of 240 days, 240 to 365 days, or more than 365 days week, 20mg / week to 60mg / week, 30mg / week to 75mg / week, or 40mg / week or in subjects with Crohn's disease or ulcerative colitis to deliver at a rate of 90 mg / week In some embodiments, the cell chamber device is maintained at a constant temperature. the period of time during which serum or plasma concentrations of the moiety are effective to achieve a therapeutic benefit in the subject; For example, at least 60 days after implantation (e.g., 60-70 days, 65-75 days, 7 0-90 days, 90-120 days, 120-240 days, 240-365 days, or 3 For a period of more than 65 days, subjects had a mean of 2.5 μg / mL to 7.5 μg / mL, 4 μg / mL to 7μg / mL, 5μg / mL~8μg / mL, 5μg / mL~10μg / mL, 5μg / mL mL~50μg / mL, 8μg / mL~15μg / mL, 10μg / mL~20μg / m L, 16μg / mL~20μg / mL, 18μg / mL~21μg / mL, 21μg / m L~30μg / mL, 25μg / mL~35μg / mL, or 31μg / mL~45μ The serum saturation level is maintained in subjects with Crohn's disease or ulcerative colitis at 0.5 mg / mL. In some embodiments, the Crohn's disease subject has moderate to severe active Crohn's disease (e.g., The patient may have a Roan Disease Activity Index (CDAI) score of 220 to 450. Treatment is Clinical responses can be achieved in patients with moderate to severe active Crohn's disease. For example, treatment may result in mucosal healing. Treatment may also involve reducing, eliminating, or reducing and eliminating the use of corticosteroids by the patient. In some embodiments, the ulcer A subject with ulcerative colitis may have moderate to severe active ulcerative colitis (e.g., 2-3 Mayo score of 6-12 with endoscopic subscore). Treatment is moderate to severe. Induction and maintenance of clinical response, clinical improvement, and clinical outcomes in patients with severe to severely active ulcerative colitis Treatment may also result in the induction and maintenance of remission or mucosal healing. This may result in the reduction, elimination, or reduction and elimination of corticosteroid use (e.g., corticosteroid use). corticosteroid-free remission).

[0198] In another aspect, provided herein is a method for treating a subject with inflammatory bowel disease (IBD). A method of treating a subject comprising administering to a subject a cell chamber device as disclosed herein. wherein the cells in the cell chamber are incubated with an anti-α4β7 antibody (e.g., vedolizumab or or abrilumab), or an antigen-binding portion thereof. Examples of inflammatory bowel diseases (IBD) that can be caused by Enterocolitis, celiac disease, non-tropical sprue, seronegative arthropathy and associated enteropathy, microscopic or collagenous colitis, eosinophilic gastroenteritis, or pouchitis occurring after rectal resection, and ileoanal anastomosis In some embodiments, the cell chamber device may include a cell chamber containing an antibody or an antigen thereof. The serum or plasma concentration of the binding moiety is determined to be effective to achieve a therapeutic benefit in the subject. For example, at least 30 days after implantation (e.g., at least 30 days, at least 4 5 days, at least 55 days, at least 60 days, at least 70 days, at least 90 days, 3 0-90 days, 40-60 days, 50-70 days, 60-70 days, 60-90 days, 6 0-120 days, 65-75 days, 70-90 days, 90-120 days, 120-24 days 0 days, 240-365 days, or more than 365 days) in subjects with at least 5 μg / mL (e.g., at least about 5 μg / mL, at least about 10 μg / mL, at least about 15 μg / mL, at least about 17 μg / mL, at least about 20 μg / mL, or In some embodiments, the IBD concentration is maintained in subjects with IBD such that the IBD concentration is greater than 20 μg / mL. In the cell chamber device, the serum or plasma concentration of the antibody, or antigen-binding portion thereof, for a period of time effective to achieve a therapeutic benefit in the subject, e.g., at least 10 days after implantation. 60 days (e.g. 60-70 days, 65-75 days, 70-90 days, 90-120 days) days, 120-240 days, 240-365 days, or more than 365 days), in the subject, .5μg / mL~7.5μg / mL, 4μg / mL~7μg / mL, 5μg / mL~8μ g / mL, 5μg / mL~10μg / mL, 5μg / mL~50μg / mL, 8μg / m L~15μg / mL, 10μg / mL~20μg / mL, 16μg / mL~20μg / m L, 18μg / mL~21μg / mL, 21μg / mL~30μg / mL, 25μg / m IBD patients should have a serum creatinine level of 31μg / mL to 35μg / mL, or 31μg / mL to 45μg / mL. The object is maintained within the object.

[0199] In some embodiments, the cell chamber device is adapted to treat Crohn's disease, ulcerative colitis, or other conditions. The drug is implanted into the peritoneum, mesenteric, or peritoneal space of a subject with some form of IBD. In embodiments, the cell chamber device is used to treat Crohn's disease, ulcerative colitis, or other forms of I. In subjects with BD, the omentum, the lesser omentum (i.e., the omental bursa or omental sac), the greater omentum, and the lesser omentum or implanted in or near the stomach, in or near the small intestine, or in or near the large intestine Be absorbed.

[0200] In one embodiment, provided herein is a method for treating a subject with primary sclerosing cholangitis. A method comprising administering to a subject a device disclosed herein. In certain such embodiments, a subject with primary sclerosing cholangitis is administered an antibody, For example, an anti-α4β7 antibody (e.g., vedolizumab or abrilumab), or its antigen In some embodiments, the cell channel is administered a device containing cells that secrete the binding moiety. The device is configured to measure the serum or plasma concentration of an antibody, or an antigen-binding portion thereof, in a subject. A period of time effective to achieve a therapeutic benefit, e.g., at least 30 days (e.g., 12 months) after implantation. For example, at least 30 days, at least 45 days, at least 55 days, at least 60 days, At least 70 days, at least 90 days, 30 to 90 days, 40 to 60 days, 50 to 70 days, 60-70 days, 60-90 days, 60-120 days, 65-75 days, 70-90 days , 90-120 days, 120-240 days, 240-365 days, or over 365 days) for a period of time, the subject receives at least 5 μg / mL (e.g., at least about 5 μg / mL, at least about 10 μg / mL, at least about 15 μg / mL, at least about 17 μg / mL, at least about 20 μg / mL, or greater than 20 μg / mL) in patients with primary sclerosing cholangiocarcinoma In some embodiments, the cell chamber device is maintained in a subject having vasculitis. or an antigen-binding portion thereof, the serum or plasma concentration of which achieves a therapeutic benefit in the subject. For example, at least 30 days (e.g., at least 30 days) after implantation. days, at least 45 days, at least 55 days, at least 60 days, at least 70 days, at least At most 90 days, 30-90 days, 40-60 days, 50-70 days, 60-70 days, 6 0-90 days, 60-120 days, 65-75 days, 70-90 days, 90-120 days , 120-240 days, 240-365 days, or more than 365 days) 2.5μg / mL~7.5μg / mL, 4μg / mL~7μg / mL, 5μg / mL~ 8μg / mL, 5μg / mL~10μg / mL, 5μg / mL~50μg / mL, 8μg / mL~15μg / mL, 10μg / mL~20μg / mL, 16μg / mL~20μg / mL, 18μg / mL~21μg / mL, 21μg / mL~30μg / mL, 25μg / mL to 35 μg / mL, or 31 μg / mL to 45 μg / mL, In some embodiments, the cell chamber device is maintained in a subject with sclerosing cholangitis. The device delivers the antibody, or antigen-binding portion thereof, to achieve a therapeutic benefit in the subject. For example, at least 60 days (e.g., 60 to 70 days) after implantation. 65-75 days, 70-90 days, 90-120 days, 120-240 days, 240 days and over 365 days or more), 10mg / week to 50mg / week, 20mg / week to 6 at rates of 0 mg / week, 30 mg / week to 75 mg / week, or 40 mg / week to 90 mg / week The drug is maintained in a subject with primary sclerosing cholangitis to deliver the drug.

[0201] In some embodiments, the cell chamber device is administered to the abdomen of a subject with primary sclerosing cholangitis. In some embodiments, the cell chamber device is implanted in the peritoneal, mesenteric, or peritoneal space. The stool was extracted from the omentum of subjects with primary sclerosing cholangitis, and the lesser omental bursa (i.e., the omental bursa or omentum) omentum), in or on the greater omentum, lesser omentum, in or near the liver, or in or near the sac is embedded in.

[0202] In one embodiment, provided herein is a method for treating lymphocytic esophagitis or eosinophilic esophagitis. A method of treating a subject having a rheumatoid arthritis, comprising administering to the subject a device disclosed herein. In certain such embodiments, the method comprises administering to a patient having eosinophilic esophagitis. The subject is receiving an antibody, e.g., an anti-α4β7 antibody (e.g., vedolizumab or abrilumab), In some embodiments, the patient is administered a device containing cells secreting the antigen-binding portion thereof. The cell chamber device is configured to detect when the serum or plasma concentration of the antibody, or antigen-binding portion thereof, is for a period of time effective to achieve a therapeutic benefit in the subject, e.g., at least 10 days after implantation. at least 30 days (e.g., at least 30 days, at least 45 days, at least 55 days, at least At least 60 days, at least 70 days, at least 90 days, 30-90 days, 40-60 days, 5 0-70 days, 60-70 days, 60-90 days, 60-120 days, 65-75 days, 70-90 days, 90-120 days, 120-240 days, 240-365 days, or at least 5 μg / mL (e.g., at least about 5 μg / mL) in a subject for a period of more than 365 days g / mL, at least about 10 μg / mL, at least about 15 μg / mL, at least about 1 7 μg / mL, at least about 20 μg / mL, or greater than 20 μg / mL) In some embodiments, the esophageal mucosa is maintained in a subject with eosinophilic esophagitis (e.g., above the diaphragm). The cell chamber device is configured to detect when the serum or plasma concentration of the antibody, or antigen-binding portion thereof, is for a period of time effective to achieve a therapeutic benefit in the subject, e.g., at least 10 days after implantation. at least 30 days (e.g., at least 30 days, at least 45 days, at least 55 days, at least At least 60 days, at least 70 days, at least 90 days, 30-90 days, 40-60 days, 5 0-70 days, 60-70 days, 60-90 days, 60-120 days, 65-75 days, 70 to 90 days, 90 to 120 days, 120 to 240 days, 240 to 365 days, or For a period of more than 365 days, subjects had a mean of 2.5 μg / mL to 7.5 μg / mL, 4 μg / mL L~7μg / mL, 5μg / mL~8μg / mL, 5μg / mL~10μg / mL, 5μ g / mL~50μg / mL, 8μg / mL~15μg / mL, 10μg / mL~20μg / mL, 16μg / mL~20μg / mL, 18μg / mL~21μg / mL, 21μg / mL~30μg / mL, 25μg / mL~35μg / mL, or 31μg / mL~4 In some embodiments, the eosinophilic esophagitis concentration is maintained at 5 μg / mL. In this embodiment, the cell chamber device is configured such that the device delivers an antibody, or antigen-binding portion thereof, to a target for a period of time effective to achieve a therapeutic benefit in, for example, at least 60 days after implantation. Days (e.g., 60-70 days, 65-75 days, 70-90 days, 90-120 days, 1 10 mg / week for a period of 20 to 240 days, 240 to 365 days, or more than 365 days 0 mg / week, 20 mg / week to 60 mg / week, 30 mg / week to 75 mg / week, or 40 mg g / week to 90 mg / week for lymphocytic or eosinophilic esophagitis is maintained within the subject having

[0203] In some embodiments, the cell chamber device is administered to the peritoneum of a subject with eosinophilic esophagitis; In some embodiments, the cell chamber device is implanted in the mesenteric or peritoneal space. In the omentum of subjects with lymphocytic esophagitis or eosinophilic esophagitis, the lesser omental bursa (i.e. , omental bursa or omental bursa), in or on the greater omentum, lesser omentum, in or near the esophagus, or diaphragm is embedded on top of the

[0204] In one embodiment, provided herein is a method for treating a subject with autoimmune hepatitis. A method comprising administering to a subject a device disclosed herein. In certain such embodiments, a subject with autoimmune hepatitis is treated with, for example, antibodies, an anti-α4β7 antibody (e.g., vedolizumab or abrilumab), or an antigen-binding portion thereof In some embodiments, the patient is administered a cell chamber device containing cells that secrete the The method further comprises determining whether the serum or plasma concentration of the antibody, or antigen-binding portion thereof, is therapeutically effective in the subject. A period of time effective to achieve benefit, e.g., at least 30 days (e.g., at least At least 30 days, at least 45 days, at least 55 days, at least 60 days, at least 7 0 days, at least 90 days, 30-90 days, 40-60 days, 50-70 days, 60 days or more 70 days, 60-90 days, 60-120 days, 65-75 days, 70-90 days, 90 days ~120 days, 120-240 days, 240-365 days, or more than 365 days) at least 5 μg / mL (e.g., at least about 5 μg / mL, at least about 10 μg / mL, at least about 15 μg / mL, at least about 17 μg / mL, at least have autoimmune hepatitis, such as a serum erythrocyte serum concentration of about 20 μg / mL or more than 20 μg / mL In some embodiments, the cell chamber device is maintained within the subject. The serum or plasma concentration of the antigen-binding moiety is effective to achieve a therapeutic benefit in the subject. period, e.g., at least 30 days after implantation (e.g., at least 30 days, at least 45 days, at least 55 days, at least 60 days, at least 70 days, at least 90 days, 30 to 90 days, 40 to 60 days, 50 to 70 days, 60 to 70 days, 60 to 90 days, 60-120 days, 65-75 days, 70-90 days, 90-120 days, 120-2 2.5 μg / m in subjects for a period of 40 days, 240 to 365 days, or more than 365 days L~7.5μg / mL, 4μg / mL~7μg / mL, 5μg / mL~8μg / mL, 5 μg / mL~10μg / mL, 5μg / mL~50μg / mL, 8μg / mL~15μg / mL, 10μg / mL~20μg / mL, 16μg / mL~20μg / mL, 18μg / mL~21μg / mL, 21μg / mL~30μg / mL, 25μg / mL~35μg / mL, or 31μg / mL to 45μg / mL, with autoimmune hepatitis In some embodiments, the cell chamber device is maintained within the subject. or an antigen-binding portion thereof for a period of time effective to achieve a therapeutic benefit in a subject, e.g., For example, at least 60 days after implantation (e.g., 60-70 days, 65-75 days, 70 90 days to 90 days, 90 days to 120 days, 120 days to 240 days, 240 days to 365 days, or 36 (more than 5 days), 10mg / week to 50mg / week, 20mg / week to 60mg / week, 30mg self-administered to deliver at a rate of 40 mg / week to 90 mg / week. maintained in subjects with immune hepatitis.

[0205] In some embodiments, the cell chamber device is administered to the peritoneum of a subject with autoimmune hepatitis; In some embodiments, the cell chamber device is implanted in the mesenteric or peritoneal space. In the omentum of a subject with autoimmune hepatitis, the lesser omental bursa (i.e., the omental bursa or omental bursa), the larger It is implanted in or on the omentum, lesser omentum, or in or near the liver.

[0206] Pathogenesis exploits the interaction of MAdCAM (e.g., MAdCAM-1) with α4β7 Diseases or pathogens may also be targeted by the antibodies delivered by the cell chamber devices described herein. α4β7 antibodies (e.g., vedolizumab or abrilumab), or antigen-binding portions thereof Examples of such diseases include those caused by the human immunodeficiency virus. Examples of immune deficiency disorders include those caused by immune deficiency syndrome (see, for example, WO2008 / 140602). See the reference.

[0207] Pancreatitis and insulin-dependent diabetes mellitus can be treated using the cell chamber device disclosed herein. an anti-α4β7 antibody (e.g., vedolizumab or abrilumab) delivered using Other diseases that can be treated with the antigen-binding portion thereof include MAdCAM (e.g., MA dCAM-1) was expressed in NOD (non-obese diabetic) mice, as well as BALB / c and SJL mice. It has been reported to be expressed by some blood vessels within the exocrine pancreas from mice. Expression of MAdCAMs (e.g., MAdCAM-1) was associated with inflammation in NOD mice. It has been reported that MAdCAM (e.g., MAdCAM-1) is induced on the endothelium in pancreatic islets. ) was the major addressin expressed by NOD islet endothelium during the early stages of insulitis ( Hanninen, A., et al., J. Clin. Invest., 92:250 9-2515(1993)). N with either anti-MAdCAM or anti-beta7 antibodies Treatment of OD mice prevented the development of diabetes (Yang et al., Diabetes es, 46:1542-1547(1997)). Furthermore, the expression of α4β7 in pancreatic islets was Accumulation of lymphocytes was observed, and MAdCAM-1 was associated with the α4β expression in mantle cell lymphoma. 7 to the blood vessels from inflamed islets (Hanninen, A., et al., J. Clin. Invest., 92:2509-2515 (1993)) or into the gastrointestinal tract was involved in the binding of lymphoma cells (Geissmann et al., Am.J. Pathol.,153:1701-1705(1998)).

[0208] Anti-α4β7 antibodies (e.g., , vedolizumab, or abrilumab), or an antigen-binding portion thereof. Other inflammatory diseases associated with the tissue include cholecystitis, cholangitis (Adams and Eks teen Nature Reviews6:244-251(2006)Grant et al., Hepatology 33: 1065-1072 (2001)), e.g. , Behçet's disease, e.g., intestinal Behçet's disease, or pericholecititis (perichondritis of the bile ducts and liver) surrounding tissues), as well as graft-versus-host disease (e.g., in the gastrointestinal tract (e.g., after bone marrow implantation) (Petrovic et al.Blood103:1542-1547(2004) Inflammation often extends beyond the mucosal surface, as seen in Crohn's disease. Therefore, chronic inflammation such as sarcoidosis, chronic gastritis, e.g., autoimmune gastritis, Sexual diseases (Katakai et al., Int. Immunol., 14:167-1 75 (2002)), and other idiopathic conditions may be amenable to treatment.

[0209] Also provided herein are methods for producing cell-cell-containing tissue using the cell chamber devices herein. The anti-α4β7 antibody (e.g., vedolizumab or abrilumab) being delivered, or its anti Further provided herein are methods for inhibiting leukocyte infiltration of mucosal tissues using the antigen-binding moiety. The proposed method is for treating cancer (e.g., α4β7-positive tumors such as lymphoma). As used herein, the antibodies delivered using the cell chamber devices herein are α4β7 antibodies (e.g., vedolizumab or abrilumab), or antigen-binding portions thereof Other examples of inflammatory diseases associated with mucosal tissue that can be treated using the method include mastitis ( mammary gland) and irritable bowel syndrome.

[0210] The cell chamber device is loaded with several cells and their ligands for α4β7 integrin. and delivering an effective amount of an anti-α4β7 antibody, or antigen-binding portion thereof, that inhibits binding to the target antigen. With respect to therapy, an effective amount is defined as the amount of the desired treatment (prophylaxis). would be sufficient to achieve the desired effect (e.g., α4β7 integrin-mediated binding and and / or in an amount sufficient to reduce or prevent signal transduction, thereby inhibiting globular adhesion and invasion and / or related cellular responses). For example, a titer sufficient to maintain saturation, e.g., neutralization, of α4β7 integrin is α4β7-associated conditions as described herein, e.g., Crohn's disease, ulcerative colitis, inflammatory bowel disease Disease, primary sclerosing cholangitis, eosinophilic esophagitis, autoimmune hepatitis, pancreatitis, cholecystitis, cholangitis, The size of the cell chamber device can induce clinical responses or remissions of tumors such as lymphoma. The number of anti-α4β7 antibody-producing cells in the lens and device is determined by measuring the amount of anti-α4β7 antibody produced in the subject. For example, anti-α4β7 antibodies can be prepared according to the methods of the present invention. The antibodies are delivered to the target cells via the cell chamber device in the amounts and for the durations disclosed herein. can be delivered to.

[0211] The dose of anti-α4β7 antibody delivered by the cell chamber device was compared with that of patients with inflammatory bowel disease. In some embodiments, the therapeutic agent may be optimized to induce clinical responses and clinical remissions in the disease. The dosing regimen altered the CD4 to CD8 ratio in the cerebrospinal fluid of treated patients. The CD4:CD8 ratio was measured in blood, lymph node aspirates, and cerebrospinal fluid (CSF). The CSF CD4+:CD8+ lymphocyte ratio in healthy individuals is typically Typically, it is about 1 or more (Svenningsson et al., J. Neurol. oimmunol.1995;63:39-46, Svenningsson et a I. Ann, Neurol. 1993;34:155-161). Immunomodulatory substances are 4:CD8 ratio can be changed to less than 1.

[0212] In one embodiment, provided herein is a method for treating rheumatoid arthritis, psoriatic arthritis, ankylosing spinal cord, or other conditions. Spondylitis, Crohn's disease, ulcerative colitis, psoriasis, hidradenitis suppurativa, uveitis, or juvenile idiopathic A method of treating a subject with arthritis, comprising administering to a subject a device as disclosed herein. In certain such embodiments, the method includes administering to a subject suffering from rheumatoid arthritis, psoriasis, or other conditions. inflammatory arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, psoriasis, hidradenitis suppurativa, uveal ulcer Subjects with idiopathic arthritis or juvenile idiopathic arthritis may be treated with, for example, an anti-TNFα antibody (e.g., Adalini et al., J. Med. Soc., 1999). antibodies such as cerebrospinal fluid (e.g., cerebrospinal fluid ... In some embodiments, the patient is administered a device containing cells that secrete an antigen-binding portion of the The chamber device contains an anti-TNFα antibody (e.g., adalimumab, certolizumab, golimumab, or infliximab), or an antigen-binding portion thereof, that provides therapeutic benefit in a subject Subjects should receive at least 20 mg (e.g., at least 20 mg, at least about 30 mg, at least about 40 mg, at least about 50 mg, at least about 60 mg, at least about 70 mg, at least about 80 mg, at least 100 mg g, at least 150 mg, at least 200 mg, at least 300 mg, or less at least 400 mg) in weekly, biweekly, monthly, or bimonthly doses It is secreted at a rate sufficient to treat rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, psoriasis, psoriatic suppurative inflammation, uveitis, or juvenile idiopathic arthritis In some embodiments, the cell chamber device is maintained in a subject having an anti-TNF antibody. alpha antibodies (e.g., adalimumab, certolizumab, golimumab, or infliximab) ), or an antigen-binding portion thereof, is present in a plasma or serum concentration that achieves a therapeutic benefit in a subject. For example, at least 30 days (e.g., at least 30 days) after implantation. days, at least 45 days, at least 55 days, at least 60 days, at least 70 days, at least At most 90 days, 30-90 days, 40-60 days, 50-70 days, 60-70 days, 6 0-90 days, 60-120 days, 65-75 days, 70-90 days, 90-120 days , 120-240 days, 240-365 days, or more than 365 days) 2.5μg / mL~7.5μg / mL, 4μg / mL~7μg / mL, 5μg / mL~ 8μg / mL, 5μg / mL~10μg / mL, 5μg / mL~50μg / mL, 8μg / mL~15μg / mL, 10μg / mL~20μg / mL, 16μg / mL~20μg / mL, 18μg / mL~21μg / mL, 21μg / mL~30μg / mL, 25μg 1μg / mL to 35μg / mL, or 31μg / mL to 45μg / mL. Uromatoid arthritis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, psoriasis, suppurative psoriasis The present invention relates to a method for treating idiopathic arthritis, including the treatment of ulcerative colitis, ... or juvenile idiopathic arthritis.

[0213] In another aspect, provided herein is a method for treating a subject with inflammatory bowel disease (IBD). A method of treating a subject comprising administering to a subject a cell chamber device as disclosed herein. The cells in the cell chamber are treated with an anti-TNFα antibody (e.g., adalimumab, certolizumab, The method of treatment according to the present invention is Examples of inflammatory bowel diseases (IBD) that can be caused by Enterocolitis, celiac disease, non-tropical sprue, seronegative arthropathy and associated enteropathy, microscopic or collagenous colitis, eosinophilic gastroenteritis, or pouchitis occurring after rectal resection, and ileoanal anastomosis In some embodiments, the inflammatory bowel disease is Crohn's disease or ulcerative colitis. is.

[0214] In another embodiment, provided herein is a method for treating ulcerative colitis or Crohn's disease. A method of treating a subject suffering from rheumatoid arthritis comprising administering to the subject a device disclosed herein. In certain such embodiments, the method comprises administering to a patient suffering from ulcerative colitis or Crohn's disease. The subject has secreted an antibody, such as an anti-integrin β7 antibody (e.g., etrolizumab). In some embodiments, the cell chamber device comprises: An anti-integrin β7 antibody (e.g., etrolizumab), or an antigen-binding portion thereof, is administered to a subject In subjects, at least 50 mg ( For example, at least about 50 mg, at least about 25 mg, at least about 75 mg, or less At least about 100 mg, at least about 125 mg, at least about 150 mg, at least about 1 75 mg, at least about 200 mg, at least about 225 mg, or at least about 25 sufficient to provide weekly, biweekly, monthly, or bimonthly doses of 0 mg maintained in subjects with ulcerative colitis or Crohn's disease so that they are secreted at a rate sufficient to In some embodiments, the cell chamber device is provided with an anti-integrin β7 antibody (e.g., etrolizumab), or an antigen-binding portion thereof, is measured in a subject A period of time effective to achieve the above benefits, e.g., at least 30 days after implantation (e.g., At least 30 days, at least 45 days, at least 55 days, at least 60 days, at least At least 70 days, at least 90 days, 30-90 days, 40-60 days, 50-70 days, 60 70 days to 70 days, 60 days to 90 days, 60 days to 120 days, 65 days to 75 days, 70 days to 90 days, 9 0-120 days, 120-240 days, 240-365 days, or over 365 days) Between subjects, 2.5μg / mL to 7.5μg / mL, 4μg / mL to 7μg / mL, 5μg / mL~8μg / mL, 5μg / mL~10μg / mL, 5μg / mL~50μg / mL, 8μg / mL~15μg / mL, 10μg / mL~20μg / mL, 16μg / mL mL~20μg / mL, 18μg / mL~21μg / mL, 21μg / mL~30μg / mL, 25 μg / mL to 35 μg / mL, or 31 μg / mL to 45 μg / mL In this manner, the compound is maintained in a subject with ulcerative colitis or Crohn's disease.

[0215] In some embodiments, the subject with ulcerative colitis or Crohn's disease is receiving anti-IL-12 / A device containing cells secreting an antibody, such as an IL-23 antibody (e.g., ustekinumab) In some embodiments, the cell chamber device is administered an anti-IL-12 / IL-23 The antibody (e.g., ustekinumab), or an antigen-binding portion thereof, provides therapeutic benefit in a subject. Subjects should receive at least 30 mg (e.g., at least about 30 mg, at least about 40 mg, at least about 75 mg, at least about 100 mg, At least about 125 mg, at least about 150 mg, at least about 175 mg, at least at least about 200 mg, at least about 225 mg, at least about 250 mg, at least about 30 0 mg, at least about 350 mg, at least about 400 mg, at least about 450 mg, at least about 500 mg, or at least about 550 mg) once a week, once every two weeks, or once a month ulcerative colitis, so that it is secreted at a rate sufficient to provide a single or bimonthly dose. In some embodiments, the cell channel is maintained in a subject with colitis or Crohn's disease. The device may contain an anti-IL-12 / IL-23 antibody (e.g., ustekinumab), or The plasma or serum concentration of the antigen-binding moiety is effective to achieve a therapeutic benefit in the subject. for a suitable period of time, e.g., at least 30 days after implantation (e.g., at least 30 days, at least At least 45 days, at least 55 days, at least 60 days, at least 70 days, at least 90 days , 30 to 90 days, 40 to 60 days, 50 to 70 days, 60 to 70 days, 60 to 90 days , 60-120 days, 65-75 days, 70-90 days, 90-120 days, 120 days or more 2.5 μg in subjects for a period of 240 days, 240 to 365 days, or more than 365 days / mL~7.5μg / mL, 4μg / mL~7μg / mL, 5μg / mL~8μg / mL , 5μg / mL~10μg / mL, 5μg / mL~50μg / mL, 8μg / mL~15 μg / mL, 10μg / mL~20μg / mL, 16μg / mL~20μg / mL, 18 μg / mL~21μg / mL, 21μg / mL~30μg / mL, 25μg / mL~35 μg / mL, or 31 μg / mL to 45 μg / mL, for ulcerative colitis or maintained in subjects with Crohn's disease.

[0216] In one embodiment, provided herein is a method for treating a subject with an autoimmune disease or cancer. A method of treatment comprising administering to a subject a device disclosed herein. In certain such embodiments, the method is directed to treating an autoimmune disease (e.g., rheumatoid arthritis) or a subject with cancer (e.g., non-Hodgkin's lymphoma, chronic lymphocytic leukemia) may receive an anti- administering a device containing cells that secrete antibodies, such as CD20 antibodies (e.g., rituximab); In some embodiments, the cell chamber device is configured to contain an anti-CD20 antibody (e.g., Ritz). The concentration of ribosomal steroids (e.g., ribosomal ximab), or an antigen-binding portion thereof, is sufficient to achieve a therapeutic benefit in the subject. at least 200 mg / m 2 (e.g., at least about 200 mg / m 2 , at least about 225 mg / m 2 , at least about 250 mg / m 2 , small At least about 300 mg / m 2 , at least about 350 mg / m 2 , at least about 400 mg / m 2 , at least about 450 mg / m 2 , at least about 500 mg / m 2 , or less Both are approximately 550 mg / m 2 ) maintained in a subject with an autoimmune disease or cancer In some embodiments, the cell chamber device is configured to contain an anti-CD20 antibody (e.g., Ritz). ximab), or an antigen-binding portion thereof, is effective in achieving a therapeutic benefit in a subject. For a sufficient period, at least 200-250 mg / m 2 , 225-275 mg / m 2 , 250-325 mg / m 2 , 300-350 mg / m 2 , 325-375 mg / m 2 , 350-400mg / m 2 , 375-425 mg / m 2 , 400-450 mg / m2 , 425-475 mg / m 2 , 450-500mg / m 2 , or 475-525 mg / m 2 Rapid enough to provide weekly, biweekly, monthly, or bimonthly doses of In some cases, the antibody is secreted at a constant rate and maintained in subjects with autoimmune disease or cancer. In embodiments, the cell chamber device is configured to contain an anti-CD20 antibody (e.g., rituximab), or The antigen-binding portion is administered to the subject for a period of time effective to achieve a therapeutic benefit in the subject. At least 150-200mg, 175-225mg, 200-250mg, 22 5~275mg, 250~325mg, 300~350mg, 325~375mg, 35 0~400mg, 375~425mg, 400~450mg, 425~475mg, 45 0~500mg, 475~525mg, 500~550mg, 525~575mg, 55 0~600mg, 575~625mg, 600~650mg, 625~675mg, 65 0~700mg, 675~725mg, 700~750mg, 725~775mg, 75 0~800mg, 775~825mg, 800~850mg, 825~875mg, 85 0~900mg, 875~925mg, 900~950mg, 925~975mg, 95 0-1000 mg, 900-1100 mg, or 1000-1200 mg once weekly, 2 Secreted at a rate sufficient to provide weekly, monthly, or bimonthly doses In some embodiments, the cells are maintained in a subject with an autoimmune disease or cancer. The cell chamber device contains an anti-CD20 antibody (e.g., rituximab), or its antigen-binding The plasma or serum concentration of the moiety is effective to achieve a therapeutic benefit in a subject, e.g., for a period of time, e.g., For example, at least 30 days after implantation (e.g., at least 30 days, at least 45 days, At least 55 days, at least 60 days, at least 70 days, at least 90 days, 30 days ~ 90 days, 40-60 days, 50-70 days, 60-70 days, 60-90 days, 60 days or more 120 days, 65-75 days, 70-90 days, 90-120 days, 120-240 days, 240 to 365 days or more) in subjects with a blood glucose level of 2.5 μg / mL to 7 .5μg / mL, 4μg / mL~7μg / mL, 5μg / mL~8μg / mL, 5μg / mL mL~10μg / mL, 5μg / mL~50μg / mL, 8μg / mL~15μg / mL , 10μg / mL~20μg / mL, 16μg / mL~20μg / mL, 18μg / mL ~21μg / mL, 21μg / mL~30μg / mL, 25μg / mL~35μg / mL or 31 μg / mL to 45 μg / mL, those with autoimmune diseases or cancer The data is maintained within the scope of the object.

[0217] In some embodiments, a subject with multiple sclerosis or Crohn's disease receives anti-alpha-4 inhibitors. The patient is administered a device containing cells that secrete antibodies, such as glioma antibodies (e.g., natalizumab). In some embodiments, the cell chamber device is configured to contain an anti-alpha-4 integrin antibody (e.g., e.g., natalizumab), or an antigen-binding portion thereof, achieves a therapeutic benefit in the subject. and administering at least 30 mg (e.g., at least about 30 mg, at least about 0.4 mg, at least about 75 mg, at least about 100 mg, at least about 125 mg, at least about 150 mg, at least about 175 mg, at least about 200 mg g, at least about 225 mg, at least about 250 mg, at least about 300 mg, or at least about 350 mg) once a week, once every two weeks, once a month, or once every two months People with multiple sclerosis or Crohn's disease In some embodiments, the cell chamber device is maintained within a subject. The plasma or serum concentration of a guinea pig antibody (e.g., natalizumab) or an antigen-binding portion thereof is for a period of time effective to achieve a therapeutic benefit in a subject, e.g., at least 30 days (e.g., at least 30 days, at least 45 days, at least 55 days, at least 60 days, at least 70 days, at least 90 days, 30-90 days, 40-60 days, 50 70 days, 60 days, 70 days, 60 days, 90 days, 60 days, 120 days, 65 days, 7 ...60 days, 120 0-90 days, 90-120 days, 120-240 days, 240-365 days, or For a period of more than 365 days, subjects had a mean of 2.5 μg / mL to 7.5 μg / mL, 4 μg / mL ~7μg / mL, 5μg / mL~8μg / mL, 5μg / mL~10μg / mL, 5μg / mL~50μg / mL, 8μg / mL~15μg / mL, 10μg / mL~20μg / mL, 16μg / mL~20μg / mL, 18μg / mL~21μg / mL, 21μg / mL mL~30μg / mL, 25μg / mL~35μg / mL, or 31μg / mL~45 The concentration of IL-1 in subjects with multiple sclerosis or Crohn's disease is maintained at 1 μg / mL.

[0218] In another embodiment, provided herein is a method for treating a subject with short bowel syndrome. The method comprises administering to a subject a device disclosed herein. In certain such embodiments, a subject with short bowel syndrome is administered a steroid hormone therapy for gastrointestinal use. The patient is administered a device containing cells that secrete a peptide therapeutic.

[0219] In some embodiments, the cell chamber device can be used to treat cancers, such as melanoma, non-small cell lung cancer, and the like. Cellular lung cancer, small cell lung cancer, lung cancer, leukemia, liver cancer, retinoblastoma, astrocytoma, glioblastoma, gingiva Cancer, tongue cancer, neuroblastoma, head cancer, neck cancer, breast cancer, pancreatic cancer, prostate cancer, kidney cancer, bone cancer, testicular cancer , ovarian cancer, mesothelioma, cervical cancer, gastrointestinal cancer, lymphoma, myeloma, brain tumors, colon cancer, sarcoma, or bladder cancer. Thus, in one embodiment, the present invention provides a method for treating a subject with urinary tract cancer. Provided herein are methods of treating a subject having cancer, comprising administering to the subject a compound or compound(s) described herein. The present invention relates to a method for treating a cancer cell comprising administering a cell chamber device as described above. In embodiments, a subject with cancer is diagnosed with a cancer-related condition that secretes a biomolecule (e.g., an antibody) that treats the cancer. For example, in some embodiments, a cell chamber device containing cells is administered. The cells in the chamber device express CD20 (e.g., rituximab), VEGF (e.g., vegetative growth factor receptors), and trastuzumab), HER2 / neu (e.g., trastuzumab), PD-L1 (e.g., abatacept) tedolizumab, avelumab, durvalumab), PD-1 (e.g., pembrolizumab, nivolumab, cemiplimab), or EGFR (e.g., cetuximab, panitumumab) In some embodiments, the cells secrete antibodies, or antigen-binding portions thereof, that specifically bind to the The cell chamber device is adapted to measure the plasma or serum concentration of the antibody, or antigen-binding portion thereof, in a subject. for a period of time effective to achieve a therapeutic benefit in the (e.g., at least 30 days, at least 45 days, at least 55 days, at least 60 days) , at least 70 days, at least 90 days, 30 to 90 days, 40 to 60 days, 50 to 7 0 days, 60-70 days, 60-90 days, 60-120 days, 65-75 days, 70 days+ 90 days, 90-120 days, 120-240 days, 240-365 days, or 365 days For a period of more than 2 days, subjects had a mean of 2.5 μg / mL to 7.5 μg / mL, 4 μg / mL to 7 μg / mL g / mL, 5μg / mL~8μg / mL, 5μg / mL~10μg / mL, 5μg / mL ~50μg / mL, 8μg / mL~15μg / mL, 10μg / mL~20μg / mL, 16μg / mL~20μg / mL, 18μg / mL~21μg / mL, 21μg / mL~ 30μg / mL, 25μg / mL~35μg / mL, or 31μg / mL~45μg / In some embodiments, the implant is maintained in the subject with cancer so that the total amount of the implant is at least 100 mL. The implant is placed subcutaneously near the site of cancer in the subject, e.g., near an organ containing a tumor. It is possible.

[0220] In some embodiments, the cell chamber device can be used to treat autoimmune diseases, e.g., transplantation. Graft-versus-host disease (GVHD), organ transplant rejection, autoimmune hepatitis, primary biliary cirrhosis, autoimmune Epidemic cholangitis, primary sclerosing cholangitis, irritable bowel syndrome (IBS), multiple sclerosis (MS), Chronic granulomatous disease, ankylosing spondylitis, scleroderma, polymyositis, (dermato)myositis, systemic vasculitis, Systemic lupus erythematosus (SLE), Crohn's disease, insulin-dependent diabetes mellitus (type 1), or ulcerative colitis. Thus, in one embodiment, Provided herein are methods of treating a subject having an autoimmune disease, comprising administering to the subject and administering the cell chamber device disclosed herein. In certain such embodiments, a subject with an autoimmune disease is administered a biological agent to treat the autoimmune disease. A cell chamber device containing cells that secrete the body molecule is administered. For example, in some embodiments In this state, the cells in the cell chamber device are transfected with α4β7 (e.g., vedolizumab, abril) tumours), CD20 (e.g., rituximab), IL-12 / IL-23 (e.g., uterine kinumab), integrin α4 (e.g., natalizumab), TNF-α (e.g., Adalimumab) tumab, certolizumab, golimumab, infliximab), integrin β7 (e.g. , etrolizumab, CD25 (e.g., basiliximab), IL-2Rα (e.g., dac antibodies that specifically bind to IgE (e.g., omalizumab), or IgE (e.g., omalizumab), or In some embodiments, the cell chamber device secretes an antigen-binding portion of an antibody or The plasma or serum concentration of the antigen-binding portion is determined to be sufficient to achieve a therapeutic benefit in a subject. for a period of time that is effective for at least 30 days after implantation (e.g., at least 30 days, At least 45 days, at least 55 days, at least 60 days, at least 70 days, at least 90 days, 30-90 days, 40-60 days, 50-70 days, 60-70 days, 60 days+ 90 days, 60-120 days, 65-75 days, 70-90 days, 90-120 days, 12 0-240 days, 240-365 days, or over 365 days) for the target period. 5μg / mL~7.5μg / mL, 4μg / mL~7μg / mL, 5μg / mL~8μg / mL, 5μg / mL~10μg / mL, 5μg / mL~50μg / mL, 8μg / mL ~15μg / mL, 10μg / mL~20μg / mL, 16μg / mL~20μg / mL , 18μg / mL~21μg / mL, 21μg / mL~30μg / mL, 25μg / mL ~35μg / mL, or 31μg / mL to 45μg / mL in autoimmune diseases In some embodiments, the cell chamber device is maintained in a subject having lymphoid tissue. It is implanted near the tissue or mesentery.

[0221] In some embodiments, the cell chamber device is used to treat cells requiring enzyme replacement therapy (ERT). In such an embodiment, the cell chamber can is defective (e.g., absent, non-functional, partially functional, or The cells may be seeded with cells expressing one or more enzymes (which may be expressed at suboptimal levels). Thus, in some embodiments, provided herein are methods for treating a subject in need of enzyme replacement therapy. A method of treatment comprising administering to a subject a cell chamber device as disclosed herein. In certain such embodiments, the device comprises: It may include cells that secrete the missing enzyme.

[0222] For example, in some embodiments, provided herein are methods for treating mucopolysaccharidosis type I (MPS I) 20. A method of treating a subject having rheumatoid arthritis, comprising administering to the subject a device disclosed herein. In certain such embodiments, the method comprises administering to a patient suffering from mucopolysaccharidosis type I (MPI). Subjects with SI) have cells that secrete enzymes such as laronidase (e.g., SEQ ID NO: 50). In certain embodiments, the device comprises a cell of SEQ ID NO: 50. an enzyme comprising the amino acid sequence of SEQ ID NO: 50, or at least 90%, 92%, 94% or more of the amino acid sequence of SEQ ID NO: 50; %, 95%, 96%, 98%, or 99% of the cells secreting the enzyme.

[0223] In other embodiments, provided herein are methods for treating patients with mucopolysaccharidosis type II (MPSII). A method of treating a subject suffering from rheumatoid arthritis comprising administering to the subject a device disclosed herein. In certain such embodiments, the method comprises administering to a patient suffering from mucopolysaccharidosis type I (MPS I) ) secrete an enzyme such as idursulfase (e.g., SEQ ID NO: 51). In one particular embodiment, the device comprises cells of SEQ ID NO:5. 1, or at least 90%, 92%, or The cells secreting the enzyme have 94%, 95%, 96%, 98%, or 99% cloning efficiency.

[0224] In other embodiments, provided herein are methods for treating metachromatic leukodystrophy (MLD). A method of treating a subject having a rheumatoid arthritis, comprising administering to the subject a device disclosed herein. In certain such embodiments, the method comprises administering to a patient suffering from mucopolysaccharidosis type 1 (MPS) I) subjects with an enzyme such as arylsulfatase A (e.g., SEQ ID NO: 52) In certain embodiments, the device comprises cells secreting the an enzyme comprising the amino acid sequence of SEQ ID NO: 52, or at least 90% of SEQ ID NO: 52; 92%, 94%, 95%, 96%, 98%, or 99% of the cells secreting the enzyme include.

[0225] In some embodiments, the cell chamber device can be used to treat lysosomal storage diseases, e.g. , Pompe disease, adult-onset glycogen storage disease II (GSD II), Gaucher disease, Fab Leigh's disease, mucopolysaccharidosis type I, mucopolysaccharidosis type II, Niemann-Pick disease (types A, B, and C) Morquio disease (including types A and B), Batten disease, Maroteaux-Lamy disease, Chromosome leukodystrophy, Tay-Sachs disease, sphingolipid disease, Hurler disease, or Subjects with Hunter syndrome can be treated. Optionally, the treated lysosomes Lysosomal storage diseases are characterized by reduced or absent activity of lysosomal enzymes in the brains of patients. Thus, in one embodiment, provided herein is a method for treating a patient with a lysosomal storage disease. A method of treating a subject, comprising administering to the subject a cell chamber device as disclosed herein. In certain such embodiments, the method comprises administering to a subject suffering from a lysosomal storage disease A subject having a lysosomal storage disease is treated with a cell channel comprising cells that secrete a biomolecule that treats the lysosomal storage disease. For example, in some embodiments, cells within a cell chamber device are administered. The cells express hexosaminidase A, alpha-galactosidase A, and glucocerebrosidase , arylsulfatase A, galactocerebrosidase, and sphingomyelinase deficient in subjects with lysosomal storage diseases, including but not limited to In some embodiments, the cell chamber device secretes an enzyme that is not present in the cell chamber device. The cells are secreting agalsidase beta, agalsidase alpha, imiglucerase, and taligravir. Sebelipase alfa, velaglucerase alfa, alglucerase, sebelipase alf α, laronidase, idursulfase, elosulfase alfa, galsulfase The enzymes secrete therapeutic agents selected from the group consisting of α-glucosidase and α-glucosidase alfa. The disclosed cell chamber device and related methods are directed to the treatment of lysosomal cells characterized by severe brain damage. In some embodiments, the cell chamber device is also useful for treating blood cell storage disorders. In another embodiment, the cell chamber device delivers biomolecules across the liquid-brain barrier. It is implanted in the elephant's brain.

[0226] In some embodiments, the cell chamber device is used to treat diabetes, thyroid disorders, or Endocrine and / or metabolic disorders such as osteoporosis can be treated. Provided herein are methods for treating a patient with an endocrine and / or metabolic disorder (e.g., diabetes). The present invention relates to a method for treating a subject suffering from a tumor, the method comprising administering to the subject a cell chamber device as disclosed herein. In certain such embodiments, the method comprises administering to an endocrine and / or or a subject with a metabolic disorder (e.g., diabetes) is treated for endocrine and / or metabolic disorders. A cell chamber device containing cells that secrete a biomolecule (e.g., insulin) is administered. For example, in some embodiments, a cell chamber device for treating diabetes may include In another embodiment, the present invention provides a method for treating thyroid disorders, comprising administering to a subject a cell therapy ... The chamber device contains cells that secrete thyroid hormones, such as levothyroxine. In an embodiment, the cell chamber device for treating osteoporosis contains RANKL( Cells secreting antibodies that specifically bind to ribosomal protein (e.g., denosumab), or antigen-binding portions thereof Includes.

[0227] In some embodiments, the cell chamber device can be used to detect and treat leukemia, lymphoma, myeloma, Blood disorders such as anemia, sickle cell anemia, or cachexia can be treated. In one embodiment, provided herein is a method of treating a subject having a blood disorder, the method comprising administering to the subject The method includes administering to the subject a cell chamber device disclosed herein. In certain such embodiments, the subject with a hematological disorder is administered a biologic agent to treat the hematological disorder. The cell chamber device containing the cells secreting the progeny is administered. For example, in some embodiments, The cells in the cell chamber device are transfected with CD20 (e.g., rituximab, obinutuzumab) ofatumumab), CD52 (e.g., alemtuzumab), CD19 (e.g., bri CD22 (e.g., inotuzumab), CD38 (e.g., daratumumab) , CD33 (e.g., gemtuzumab), or SLAMF7 (e.g., elotuzumab) They secrete antibodies, or antigen-binding portions thereof, that specifically bind.

[0228] In some embodiments, the cell chamber device is used to treat anemia, e.g., chronic kidney disease or or anemia associated with end-stage renal disease. In the present study, a subject with anemia is treated with recombinant erythropoietin (e.g., epoetin alfa, dapagliflozin, or dapagliflozin). Patients are administered a cell chamber device containing cells secreting erythropoietin alfa.

[0229] In some embodiments, the cell chamber device can be used to treat congestive heart failure, high blood pressure, heart failure, and other conditions. Cardiovascular diseases such as myopathy, myocarditis, atherosclerosis, chronic venous disease, or cardiac arrhythmias In one embodiment, provided herein are methods for treating cardiovascular disease. A method of treating a subject having a tumor comprising administering to the subject a cell chamber device as disclosed herein. In certain such embodiments, the method comprises administering a The subject has a cell chamber containing cells secreting a known biomolecule that treats cardiovascular disease. The device is administered.

[0230] In some embodiments, the cell chamber device can be used to treat atopic asthma, non-atopic asthma, Asthma, emphysema, bronchitis, chronic obstructive pulmonary disease, sinusitis, allergic rhinitis, fibrotic lung respiratory diseases such as pulmonary vascular disease, ARDS, pulmonary vascular disease / pulmonary hypertension, cor pulmonale, or cystic fibrosis In one embodiment, provided herein are methods for treating respiratory disorders. A method of treating a subject having a tumor comprising administering to the subject a cell chamber device as disclosed herein. In certain such embodiments, the method comprises administering a The subject is a cell chamber device containing cells that secrete biomolecules that treat respiratory disorders. For example, in some embodiments, the cells in the cell chamber device are administered IL-4 α (e.g., dupilumab), IL-5 (e.g., mepolizumab), IgE (e.g., o malizumab), IL-5 (e.g., reslizumab), or IL-5Rα (e.g., Ben The antibody specifically binds to the antibody, or an antigen-binding portion thereof, of the compound (ralizumab).

[0231] In some embodiments, sustained clinical remission, e.g., for a period of 6 months or 1 year after initiation of treatment. sustained through at least two, at least three, at least four visits with a caregiver within The clinical remissions reported herein can be achieved after treatment with the cell chamber devices. In some embodiments, a sustained clinical response, e.g., at least 6 months after initiation of treatment, at least Clinical responses sustained for at least 9 months, and at least 1 year, are reported in accordance with the cell chamber method described herein. This can be achieved after treatment with the device.

[0232] The present disclosure is further illustrated by the following examples. The examples provided are for illustrative purposes only. are for illustrative purposes only and are not intended to limit the scope or content of this disclosure in any way. should not be interpreted as [Example]

[0233] Example 1: Fabrication of electrospun cell chambers To construct a cell chamber device using electrospun polymer, do the following: This exemplary cell chamber device is comprised of three nanofibrous electrospun tubings. The following method involves the addition of an additional layer of polymer to the membrane. The electrospun polymer layer, the less electrospun polymer layer, or the different electrospun polymer layers It can be adapted to fabricate devices containing Trospan polymer layer(s). The method can also be used to fabricate devices lacking a central polyester film, if desired. can be adapted to.

[0234] Polyethylene terephthalate (PET) and polybutylene terephthalate (PBT) A first broom containing hexafluoroisoproposal (HFIP) Acid vials were prepared. Polyurethane (PU) chips (polycarbonate, polyether A second borosilicate binder containing HFIP (either polyethylene or polyester) was used. The vial was rotated at a speed of 45 RPM or higher until the chips were dissolved. Placed it on the child.

[0235] The PET-PBT solution was loaded into a 10 mL syringe and inserted into the pump of the Espin unit. A polyester film containing pores of 0.2 μm to 2 μm was fixed to the mandrel. The size of the mandrel and / or film was adjusted based on the desired size of the scaffold. The nanofibrous electrospun PET-PBT layer can be adjusted to 10-20 cm. m e-spinning distance, 20-23 kV e-spinning voltage, and 15-150 RPM A rotation speed was used to apply to the film.

[0236] Remove the film from the mandrel and place it with the uncoated side facing outwards. The nanofibrous electrospun PET-PBT layer was then placed over the film coating. The syringe loaded with PU solution was connected to the Espin unit. On one side of the film, a layer of nanofibrous electrospun PU was placed on the other side of the nPET-PBT layer. In this way, a three-layer scaffold containing a central film / membrane (nPET-PB The mandrel was then attached to the ethanol. The scaffolds were then rinsed with distilled water and dried. Then, remove it from the mandrel.

[0237] The scaffolds were cut into 50 mm x 40 mm sheets. The size of the scaffolds depended on the desired size of the cell chambers. Depending on the size of the scaffold, the nPU layer can be increased or decreased. The cell chamber was then folded in half to form the inner surface of the cell chamber. Three edges were secured with Sonobond The remaining edges were ultrasonically welded using a SeamMaster unit. Cut the scaffolding diagonally from the center point of the open edge to a point below each closed edge. The resulting edge was cut to a sharp point. The cell chamber was then cut into five closed sections. The chamber was closed by ultrasonic welding, with the chamber having a flat surface. After loading, make a small opening in the cavity that can serve as a cell port. The scaffold was treated with ethyldiamine to impart a positive charge to the polymer. The polymer may be immersed in water or treated with sodium hydroxide to impart a negative charge to the polymer. To reduce inflammation due to implantation trauma, a cell scaffold device was prepared as described in Example 8. The drug may optionally be loaded with tacrolimus, as described above.

[0238] Example 2: Development of Vedolizumab / Luc-ARPE-19 cells In the following examples, ARPE-19 cells were loaded into the cell chamber device. In this example, ARPE-19 cells were engineered to express vegetatively. Dolizumab is secreted (e.g., delivered to a subject when loaded into a cell chamber device) Cells can also be engineered to express luciferase for in vitro and in vivo studies. This allows for in vivo cell monitoring.

[0239] Two expression constructs were introduced into ARPE-19 cells. One expression construct was vedolizumab. The second expression construct encodes luciferase and the vedolizumab light chain. Both the light and heavy chains of vedolizumab bind to human elongation factor-1 alpha (EF-1a). Vedolizumab was expressed (without luciferase) under the control of a constitutive promoter. Fresh clones were also engineered. The cells were then grown and analyzed for levels of vedolizumab expression. (See Table 1). [Table 1]

[0240] A vedolizumab-ARPE19 clone expressing approximately 40 picograms / cell / day was isolated. Furthermore, AREP19 cells expressing both vedolizumab and luciferase were generated using the method of the present invention. Vedolizumab / Luc- ARPE19 clone).

[0241] Vedolizumab secretion per cell per day and per cm of scaffold 2 Estimated winning details To estimate the plasma concentration levels of vedolizumab that may be achieved over time based on cell counts A simulated model was prepared with 625,000 cells / cm on the scaffold. 2 estimated to grow 625,000 cells / cm 2 but 45 picograms of vedolizumab per day When secreted, approximately 28 μg of vedolizumab / cm per day 2 will be secreted. Therefore, an 8 x 10 cm device secretes approximately 4.5 mg of vedolizumab per day. As shown in the simulation, such devices are biocompatible and can be used within 60 days of implantation. Therapeutic plasma concentrations of dolizumab (17 μg / mL) would be predicted to be achieved.

[0242] Example 3: In vitro cell seeding assay with vedolizumab / Luc-ARPE-19 cells stomach ARPE-19 cells expressing vedolizumab and luciferase (vedolizumab / Lu c-ARPE19 cells) were cultured in polyethylene terephthalate-polybutylene terephthalate (PET) media. A 16mm disc consisting of an outer layer of (nPET-PBT) and an inner layer of polyurethane (PU). The seeds were sown in the field.

[0243] To assess whether cells could grow on the inner PU layer of the scaffold, Mab / Luc-ARPE19 cells were stained with CellTracker Orange (5 μM). 250,000 pre-stained cells were plated onto a 16 mm diameter plate of nPET-PBT / PU. The discs were then imaged by fluorescence imaging 24 hours after seeding. As shown in Figure 3A, cells grew on the PU side and had a monolayer appearance. No cells were detected on the outer nPET-PBT side of the scaffold. No cell migration to the PET side was observed.

[0244] The number of cells loaded on the nPET-PBT / PU discs was determined by the number of cells per 1000 cells per 1000 cells in tissue culture (TC) plates. The number of cells that can be loaded onto tissue culture plates and nPET-PBT / PU plates was compared. Images of cells on disk are shown in Figure 3B. The top row shows the C in the cytoplasm of cells in each state. The bottom row shows ellTracker Orange fluorescence, and the bottom row shows Hoechst33342 The nuclei stained with .PI. in the nPET-PBT / PU disc compared to the tissue culture plate. The number of cells in each condition was determined by quantifying the amount of DNA isolated from cells in each condition. As shown in Figure 3C, the ATP-dependent ATPase activity was measured by ELISA using 100% ATP-dependent ATPase activity assay. The amount of DNA isolated was five times that isolated from cells on tissue culture plates. At least five times more cells can be cultured on nPET-PBT / PU than on PET. Furthermore, the amount of vedolizumab secreted in each state was compared with that in nPET-PBT. Cells on PU membranes secreted five times more than cells on tissue culture plates ( Figure 3D).

[0245] Next, vedolizumab / Luc-ARPE19 cells were seeded and the cells were then used to Secretion was evaluated on nPET-PBT with charged surface modification. The disc containing the positive PET was treated with liquid ethylenediamine (nPET-PBT(EDA)). A positively charged surface was produced or treated with liquid sodium hydroxide to produce a negatively charged surface. The surface (nPET-PBT(HYD)) was generated. IgE / luciferase was administered at a density of 1 million cells / 16 mm disc using nPET. -PBT / PU, nPET-PBT, nPET-PBT(EDA), or nPET-P The cells were seeded on a disc containing BT(HYD). The cells were incubated in an incubator at 37°C. Maintain in DMEM / F-12 medium supplemented with 10% FBS under 5% CO2 conditions. The medium was replaced with fresh medium every 3–4 days. Three weeks after seeding, vedolizumab secretion (Figure 4A) was observed. ) and luminescence intensity (Figure 4B) were measured by vedolizumab ELISA and IVIS imaging, respectively. As shown in Figures 4A and 4B, vedolizumab / Luc-ARPE-1 9 cells were seeded on the surface-modified material, and the cells were able to sustain vedolizumab secretion. came.

[0246] Subsequently, they were grown on charge-modified or unmodified nPET-PBT. The cytokine secretion profile of vedolizumab / Luc-ARPE19 cells was evaluated. nPET-PBT / PU, nPET-PBT, nPET-PBT(EDA), or nP Vedolizumab / Luc-ARPE19 cells on discs containing ET-PBT(HYD). Three weeks after seeding, cytokine production by vedolizumab / Luc-ARPE19 cells was The data were collected from cells seeded on nPET-PBT / PU membranes. The secretion data were normalized by the secretion data from ARPE-19 cells as shown in Table 2. The secretion of kinesin only changed 0.5-1.2 fold, indicating that the charged surface modification affected cell function. It showed that it did not. [Table 2]

[0247] Example 4: Vedolizumab / Luc-ARP in a nanofibrous polymer cell chamber device In vitro and in vivo cellular distribution of E-19 cells In this study, two cell chamber devices were tested. The first device was the nPET The scaffold contained an outer layer of PBT and an inner layer of PU surrounding the cell chamber. The device is shown in Figure 1C. Other devices have the same nPET-PBT / PU scaffold, but with the addition of The cell chamber contains nanofibrous polybutylene (nPBT) modified with pores. The device contained a second scaffold attached to it. This device is shown in Figure ID. To assess the distribution of cells in vitro, the cell chamber was filled with vedolizumab / Luc- ARPE19 cells were loaded. Cell distribution was assessed by luminescence imaging 4 days after cell seeding. As shown in Figure 5A, cells were able to invade both devices, i.e., with the inner nPBT scaffold. The cells were distributed on the nPET-PBT / PU cell chamber with or without PBS.

[0248] nPET-PBT with or without inner nPBT scaffold after in vivo implantation To assess cell viability and distribution on the PU / BALB chamber, both chambers were The mice were implanted (subcutaneously, near the shoulder) into 1 / c nude mice. The mice were then analyzed by luminescence imaging and blood sampling. The cells were incubated with D-luciferin / PBS and evaluated twice a week over a 62-day period. The solution (15 mg / mL, 100 μL / 10 g) was injected intraperitoneally, followed by 10 min postinjection. The results were analyzed by imaging mice at 100°C for 10 min at 4°C. As shown in Figures 5B and 5C, which show the luminescence intensity from the chamber, the cells survived for up to 65 days. Cell channels were cultured with or without an inner nPBT scaffold for 90 days or 90 days, respectively. Furthermore, as shown in Figure 5D, vedolizumab was viable within the implanted device. At 90 days after implantation, Western blot analysis was performed on mice implanted with either chamber device. Blot (primary antibody: goat anti-human antibody, Fc region specific; secondary antibody: rabbit anti-goat antibody, Detected by HRP conjugate).

[0249] To assess whether cells leaked out of the chamber, D-luciferin was added to the cell chamber. Mice implanted with the chamber were injected intraperitoneally, and IVIS imaging was performed 8 minutes after injection. Then, the mouse was euthanized, the chamber was removed, and IVIS imaging was performed again to confirm that the cells The mouse was positioned on its back. As shown in Figure 5E, the chamber After removal, there was no detectable luminescent signal on the back of the mouse, indicating that vedolizumab / Luc -ARPE19 cells were not leaking from the chamber. 42 days after implantation The removed and explanted chambers were evaluated. As shown in Table 3, the explanted chambers In addition, as shown in Figures 5F and 5G, significant No significant fibrotic response was visually observed. [Table 3]

[0250] To evaluate the inflammatory response to various scaffold materials over time, mice were treated with: (i) One side is electrospun nPET-PBT and the other side is electrospun nPU. Coated solid (non-nanofiber) PET sheet with 0.4 μm pores. or (ii) a 0.4 μm scaffold not coated with nanofibrous polymer. The scaffolds containing either a solid PET sheet with micropores were implanted. 41 days after implantation The scaffold was maintained in the mouse, and the histology of the area surrounding the implant was analyzed by H&E staining. Figure 5H (left panel) shows the results of the coating of nPET-PBT and nPU. The area surrounding the first scaffold containing the solid PET sheet is shown in Figure 5H (right panel). ) a second scaffold containing a solid PET sheet without a nanofibrous coating was taken. As shown in Figure 5H, the inflammatory response to the nanofibrous scaffold material was The fibrous polymer coating was significantly reduced compared to scaffolds lacking the coating.

[0251] Example 5: In vitro cell attachment assay with vedolizumab / Luc-ARPE-19 cells stomach To assess cell attachment on the surface-modified materials, cells were cultured to express luciferase. The engineered ARPE-19 cells (Luc-ARPE-19 cells) were then subjected to nPET-PB T, nPET-PBT(EDA), or nPET-PBT(HYD) discs Cell adhesion to normal tissue culture (TC) plates was evaluated as a control. 400,000 cells were seeded per disc of material, and the adherent cells were The percentage of cells was assessed at multiple time points after seeding.

[0252] As shown in Figure 6, ARPE-19 cells express all three endogenous endothelial cells in a time-dependent manner. Charged materials adhered rapidly to the Lectrospan surface. All materials showed cell adhesion and loaded 50-60% of the cells within 60 minutes. The charged material (nPET-PBT(EDA) or nPET-PBT(HYD)) is 180 After 2 min of incubation, the cell load was greater than that of unmodified nPET-PBT. The cells were packed (90% adherent cells).

[0253] These results suggest that cells rapidly attached to the electrospun materials in a time-dependent manner. In addition, the charged surface modification improved the amount of cell loading over time. Due to the short time required for attachment, the cell chambers can be loaded with cells on the day of implantation. can.

[0254] Example 6: In vivo implantation of vedolizumab / Luc-ARPE-19 cells Sei nPET-P to test cell retention on electrospun scaffolds in vivo Consists of BT, nPET-PBT(EDA), or nPET-PBT(HYD) A 16 mm membrane disk was coated with vedolizumab and luciferase (vedolizumab / Luc-A Stably produce RPE19 cells (400,000 cells per 16 mm disc). The cells expressing the IL-1 receptor were seeded and implanted subcutaneously into female nude mice. The discs used in the experiment do not contain enclosed chambers. Administration of vedolizumab / Luc-ARPE19 cells was evaluated as a control. Each disc was loaded on the day of loading. Scaffolds were implanted subcutaneously 24 or 4 hours after loading. The mice were administered the drug by intravenous infusion (n = 4 mice). Luminescence was monitored by live cell imaging every 3-4 days. The cells were then cultured and monitored over a 70-day period to determine the extent of cell retention. As shown, ARPE-19 cells on electrospun materials were significantly higher than cells injected subcutaneously. The retention time at the implant site was longer than that of the control group.

[0255] Example 7: Vedolizumab / Luc-ARP on tacrolimus-treated nPET-PBT In vitro cell seeding assay with E-19 cells To reduce inflammation caused by implantation trauma, the cell scaffold device was treated with tacrolimus (FK5 06) can be loaded. Cell seeding and biomolecules on tacrolimus-treated materials Electrospun nPET-PBT to assess secretion Before the procedure, the polymer solution was diluted with 0% tacrolimus, 2% tacrolimus, or 4% tacrolimus. The tablets were loaded with tacrolimus by dissolving the mussels.

[0256] Incorporation of tacrolimus into nPET-PBT was confirmed by electrospun tacrolimus from the material. The extracted solution was then analyzed using a high performance liquid crystal display (HPLC). The results were analyzed by high performance liquid chromatography (HPLC). As shown in Figure B (Tacrolimus after extraction), HPLC analysis confirmed that the electrospun nP It was shown that there was no drug degradation after loading of tacrolimus onto ET-PBT.

[0257] To evaluate the bioactivity of tacrolimus-loaded nPET-PBT, T cells were Incubation in culture medium exposed to crolimus-loaded nPET-PBT, in vitro A T cell activation assay was performed. Electrospun nPET-PBT was prepared in 10% PBS. The cells were immersed in 400 μL of RPMI 1640 supplemented with fetal bovine serum (FBS) and incubated for 24 hours. The nPET-PBT was then added to fresh pre-warmed medium. The nPET-PBT was transferred to a pre-warmed fresh PBS and incubated for 24 hours (second sample). The cells were then transferred to fresh medium and incubated for 24 hours (third sample). Human PBMCs were collected for drug concentration assessment by ISA / HPLC. The samples were incubated for 30 minutes. Subsequently, human PBMCs were incubated with CD3 / CD28 activated The T cells were activated with Dynabeads for 48 hours. Then, the supernatant from the T cell culture was isolated. , and tested for IL-1β production. As shown in Figure 8C, tacrolimus-loaded Culture media exposed to nPET-PBT inhibited T cell proliferation, as measured by IL-1β levels. This inhibits the activation of tacrolimus, which remains effective after it is loaded into the electrospun material. The results show that tacrolimus was gradually released over a 3-day period (FIG. 8C).

[0258] Next, in vitro assays were performed to confirm the efficacy of tacrolimus-loaded electrospun nPs. ET-PBT inhibits cell growth and vedolizumab production by vedolizumab / Luc-ARPE19 cells. We evaluated whether the administration of tacrolimus or not affected the secretion of tacrolimus. In PET-PBT, vedolizumab / Luc-ARPE19 cells (200,000 cells / The cells were seeded on tacrolimus-loaded nPET-PBT discs for 3 days. After growth, the surface of the nPET-PBT material was visually inspected by fluorescent imaging. As shown in Figure 8D, tacrolimus had no detectable effect on cell proliferation or growth. The cells were further evaluated 3 weeks after seeding. As shown, vedolizumab / Luc-ARPE-19 cells express tacrolimus-loaded nP When plated on ET-PBT, the cells were able to sustain vedolizumab secretion. The results showed that tacrolimus was an off-target inhibitor of vedolizumab / Luc-ARPE19 cells. This indicates that the compound had no effect on the efficacy of the compound.

[0259] Subsequently, the beds grown on nPET-PBT with or without tacrolimus were analyzed. The cytokine secretion profile of lizumab / Luc-ARPE19 cells was evaluated. , grown over 3 days on tacrolimus-treated nPET-PBT, as described above. Three weeks after seeding, cytokine secretion was assessed for cells seeded on each material. Data were collected from cells seeded on nPET-PBT membranes without tacrolimus treatment. The secretion data were normalized by the secretion data. As shown in Table 4, the secretion data from ARPE-19 cells There was only a 0.5- to 2-fold change in cytokine secretion, suggesting that tacrolimus does not affect cell function. It showed that there was no [Table 4]

[0260] Example 8: Adalimumab / ARPE-19 cells and ustekinumab / ARPE-19 cells In vitro cell seeding assay using In the same manner as described above for the production of ARPE-19 cells secreting vedolizumab, Adalimumab (adalimumab / ARPE19 cells) or ustekinumab (ustekinumab) ARPE-19 cells were generated to express and secrete the IgG1-dependent cloning factor (IgG / ARPE19 cells).

[0261] Adalimumab / ARPE19 cells or ustekinumab / ARPE19 cells were treated with poly(A) an outer layer of polybutylene terephthalate-polybutylene terephthalate (nPET-PBT), and The seeds were seeded onto 16 mm discs containing an inner layer of polyurethane (PU).

[0262] Biomolecules (i.e., adalimumab) loaded onto nPET-PBT / PU cells or ustekinumab) secretion was compared with the number of cells that could be loaded onto tissue culture (TC) plates. As shown in Figures 9A and 9B, cells on the nPET-PBT / PU membrane were More adalimumab (Figure 9A) or ustekinumab (Figure 9B) than cells on the woven culture plate. 9B).

[0263] Example 9: Functional activity of antibodies secreted using the cell chamber device The functional activity of antibodies secreted by ARPE-19 cells was determined by in vitro assays. Adalimumab, ustekinumab, or vedolizumab (adalimumab-AR) PE-19, ustekinumab-ARPE-19, and vedolizumab-ARPE-19) ARPE-19 cells secreting IgG were cultured in SFM4 MegaVir medium at a density of 50,000 cells. The cells were seeded in cell culture flasks at a density of 100 cells / cm^2 and cultured for 15 days. The amount of each antibody administered was determined for adalimumab ELISA, ustekinumab ELISA, and bedrin. The results were determined based on the ELISA assay for IgE (see Tables 1 and 5). As outlined, cell culture medium containing antibodies secreted by the cells is subjected to antibody functional activity. The assay was carried out.

[0264] Adalimumab is an anti-TNFα antibody. To assay adalimumab activity in culture medium, L-929 cells were cultured in 10% F Culture 96-well cells at a density of 5,000 cells / well in DMEM supplemented with BS. After 20 hours of incubation, the cells were treated with actinomycin D (2 The cells were then sensitized with human TNFα (1 ng / mL) for 2 hours. Incubate the cells with culture medium containing antibodies secreted by RPE-19 cells. After a further 20 hours of incubation, the cells were analyzed by CCK-8 assay. Viability was assessed and tested for neutralization of human TNFα activity. Thus, these results suggest that adalimumab secreted by ARPE-19 cells is essential for T This shows that the NFα neutralizing activity was maintained.

[0265] Vedolizumab is a humanized α4β7 integrin antibody. RPE-19 clone and one vedolizumab / luciferase-expressing ARPE-19 clone To assay for vedolizumab activity in cell culture media from different Using chromatometry, we identified α4β7 integrin-specific HuT-78 cells. The α4β7 integrin binding assay was performed using the cells in SFM4MegaVir medium. These medium samples were cultured at 4°C for 2 weeks, and the vedolizumab concentration was measured by vedolizumab ELISA. The binding activity of the conditioned medium was determined by the amount of the antibody secreted from CHO cells. Competitive assay using fluorescently labeled purified vedolizumab (R-PE labeled MLN002) As shown in Figure 10B, the secreted ATP by ARPE-19 cells was Vedolizumab, which is a marker for MLN002, inhibited the binding of R-PE-labeled MLN002. showed that vedolizumab secreted by ARPE-19 cells is functional and binds to α4β7 inactivators. 1 shows that the antibody bound to tegrin and CHO cell-derived vedolizumab. [Table 5]

[0266] Example 10: Vedolizumab on nPET-PBT / PET / nPU cell chamber device In vitro expression assay using / Luc-ARPE-19 cells In this example, AR cells with different densities stably expressing vedolizumab and luciferase were cultured. In vitro production of vedolizumab in a cell chamber device seeded with PE-19 cells The cell chamber device consisted of an outer layer of nPET-PBT, an inner layer of nPU, and A porous, non-nanofiber PET membrane (0.4 μm pore size) between the inner and outer layers. The field contained nPET-PBT (as shown in Figures 1E-1G and described further below). / PET / nPU cell chamber device). For comparison, the Theracyte immunoisolation A separation device was evaluated that did not contain nanofiber material, but instead contained a woven polyester outer layer. layer, a vascularized membrane (with 5 μm PTFE pores), a nonwoven polyester insert, and The Theracyte device contains an immunoisolation membrane with 0.45 μm PTFE pores. For example, Rafeal, et al., (1999). European Surg ical research, 31(3), 249-258.

[0267] Nanofibers to generate nPET-PBT / PET / nPU cell chamber devices The polymer membranes were engineered by electrospinning (e.g., Huang, et al. .Composites Science and Technology(2003) 63:2223-2253) and assembled by ultrasonic welding. Resistant polyethylene terephthalate (PET) and polybutylene terephthalate (PBT) nPET-PBT) outer layer, a commercial non-fibrous PET membrane with a central 0.4 μm pore, and an inner inner layer containing electrospun nanofibrous polyurethane (nPU) polymer. The scaffold consisted of a trilaminate (nPET-PBT / PET / PU) containing a layer of nPE. Cutting a flat sheet of T-PBT / PET / nPU and applying ultrasonic cutting / sealing process After assembly, an air leak test was performed as a quality control. For ease of use, a blunt needle (23G) is inserted into the loading port and the device is placed in an individual sterile packet. The structure of the nPET-PBT / PET / PU device was shown in Figure 1. The SEM image of each membrane in the device is shown in Figure 1F, visualized by H&E staining. An image of the chamber 44 days after cell loading is shown in Figure 1G.

[0268] ARPE-19 cells stably expressing vedolizumab and luciferase were cultured in three different The particles were loaded into the nPET-PBT / PET / nPU chamber at densities of 2.5 million, 5 million, and 10 million. , and 10 million cells / chamber. The day after cell loading, the chamber was filled with fresh Megavi The cells were incubated in rSFM for 2 hours, followed by reconstitution of the chamber with 150 μg / ml D-luciferin. By placing it on a medium containing fetal fetal grafts, IVIS Spectrum Imaging Optical imaging was performed using the ng platform (PerkinElmer). The luminescence image of the cell chamber in Figure 11A is shown. The data were taken at constant intervals around the device. Liv by delimiting the cardiac region of interest (ROI) and quantifying total radiance in photons / second. The fluorescence intensity was analyzed using Ing Image software (PerkinElmer). The linearity of the concentration of vedolizumab in the medium versus the cell number was evaluated (Figure 11B). The linearity of vedolizumab secretion versus cell number was assessed by vedolizumab ELISA assay. This data indicates that ARPE-19 cells cultured in the cell chamber When the cells were incubated, they expressed detectable levels of luminescence, actively transferring vedolizumab from the chamber into the medium. Demonstrate secretion.

[0269] Example 11: In vivo production of vedolizumab in a cell chamber device In this example, in vivo production of vedolizumab was demonstrated using cell cultures implanted in immunodeficient mice. The cell chamber device was evaluated in the cell chamber device shown in Figures 1E to 1G and Example 10. An outer layer of nPET-PBT and an inner layer of nPU, as further described, and between the inner and outer layers. The porous, non-nanofibrillar PET membrane (0.4 μm pore size) and the scaffold (n PET-PBT / PET / nPU cell chamber device). The Theracyte immunoisolation device was also evaluated, as described in.

[0270] Vedolizumab and / or luciferase from each cell chamber device in vivo To evaluate cell viability and production of β-lactamase, nPET-PBT / PET / nPU cell chambers were used. Vedolizumab / Lucifera in the Vasa device and Theracyte cell chamber device T cells were loaded with either vedolizumab- or ARPE19 cells expressing ribozyme. The mice were then subcutaneously implanted into immunodeficient mice (i.e., nude mice) lacking the antibody. Cell viability within the cell chamber device was assessed by imaging. using the ectrum Imaging platform (PerkinElmer) In vivo imaging was performed to detect luminescence. Mice were intraperitoneally injected with D-luciferin. 10 minutes after injection, the mice were anesthetized with isoflurane. Bottom: Sequential scans were performed in the imaging chamber at 1-minute intervals for 30 minutes. PBT / PET / nPU cell chamber (top row) or Theracyte device (bottom row) ) and nude mice implanted with luminescence images taken at peak signal. As shown in Figure 12A, the cells were viable and remained in the implanted portion of the cell chamber device. It remained localized at the site.

[0271] The level of luminescence in each mouse was measured in a fixed region of interest (R...

Claims

1. 1. A device comprising a multi-layer scaffold surrounding a cell chamber, said multi-layer scaffold comprising an outer layer and an inner layer in contact with the cell chamber, the outer layer and the inner layer each comprising a nanofibrous polymer The device includes a reflector.

2. The outer layer is made of nanofibrous polyethylene terephthalate and polybutylene terephthalate. The device of claim 1 , comprising:

3. The outer layer is made of electrospun polyethylene terephthalate and polybutylene terephthalate. The device of claim 1 , further comprising a rate.

4. The device according to any one of claims 1 to 3, wherein the inner layer comprises nanofibrous polyurethane. Vice.

5. The device of claim 4 , wherein the inner layer comprises electrospun polyurethane.

6. The inner layer is made of nanofibrous polyethylene terephthalate and polybutylene terephthalate. The device according to any one of claims 1 to 3, comprising:

7. The inner layer is made of electrospun polyethylene terephthalate and polybutylene terephthalate. The device according to any one of claims 1 to 3, comprising a rate.

8. 10. The method of claim 9, wherein the outer layer and / or the inner layer comprises one or more charged surface modifications.

10. The device according to claim 1.

9. The device of claim 8 , wherein the outer layer and / or the inner layer have a net positive charge. 。

10. The device of claim 9 , wherein the scaffold is treated with ethylenediamine.

11. The device of claim 8 , wherein the outer layer and / or the inner layer have a net negative charge. 。

12. The device of claim 11 , wherein the scaffold is treated with sodium hydroxide.

13. 10. The method of claim 1, wherein the outer layer and / or the inner layer comprises an anti-inflammatory agent. device.

14. The device of claim 13 , wherein the anti-inflammatory agent is a calcineurin inhibitor.

15. The device of claim 14, wherein the anti-inflammatory agent is tacrolimus.

16. The device of claim 13 , wherein the anti-inflammatory agent is a pyridone.

17. 17. The device of claim 16, wherein the anti-inflammatory agent is pirfenidone.

18. The device of claim 13 , wherein the anti-inflammatory agent is a phosphodiesterase inhibitor.

19. 19. The device of claim 18, wherein the anti-inflammatory agent is roflumilast.

20. 10. The device according to any one of the preceding claims, wherein the outer layer and / or the inner layer comprises pores. Vice.

21. 21. The device of claim 20, wherein the pores are sized to allow the passage of biomolecules. Vice.

22. 22. The device of claim 21, wherein the biomolecule is 250 kDa or less.

23. the pores are sized to allow passage of an antibody, or an antigen-binding portion thereof; 21. The device of claim 20.

24. 24. Any of claims 20 to 23, wherein the pores are sized to prevent the passage of cells.

2. The device according to claim 1.

25. The pores allow cells on one side of the multilayer scaffold to contact cells on the other side of the multilayer scaffold. A device according to any one of claims 20 to 24, sized to prevent vinegar.

26. The device according to any one of claims 20 to 25, wherein the pores have a diameter of 1 μm or less. Vice.

27. 27. The method according to claim 20, wherein the pores have a diameter of 0.5 μm or less. device.

28. The multi-layer scaffold further comprises a porous membrane positioned between the inner layer and the outer layer.

10. A device according to any one of the preceding claims.

29. 30. The device of claim 28, wherein the porous membrane comprises polyethylene terephthalate.

30. 30. The method of claim 29, wherein the porous membrane comprises nanofibrous polyethylene terephthalate. device.

31. 30. The nanoporous membrane of claim 29, wherein the nanoporous membrane comprises non-nanofiberous polyethylene terephthalate. device.

32. 10. The method of claim 9, wherein the porous membrane comprises pores sized to allow passage of the biomolecules.

32. A device according to any one of claims 28 to 31.

33. 33. The device of claim 32, wherein the biomolecule is 250 kDa or less.

34. The membrane pores are sized to allow passage of the antibody, or antigen-binding portion thereof. The device according to any one of claims 28 to 33.

35. 35. The method of claim 28, wherein the membrane pores are sized to prevent the passage of the cells.

10. The device according to claim 1.

36. The membrane pores allow cells on one side of the multilayer scaffold to contact cells on the other side of the multilayer scaffold. A device according to any one of claims 28 to 35, sized to prevent vinegar.

37. 1. A device comprising a multi-layer scaffold surrounding a cell chamber, said multi-layer scaffold comprising: (i) Nanofibrous polyethylene terephthalate and polybutylene terephthalate An outer layer; (ii) a non-nanofibrillar membrane positioned between an inner layer and the outer layer, the membrane comprising: the non-nanofibrillar membrane comprising nanopores sized to prevent passage of cells across the membrane. A membrane and (iii) an inner layer comprising nanofibrous polyurethane.

38. 1. A device comprising a multi-layer scaffold surrounding a cell chamber, said multi-layer scaffold comprising: (i) Nanofibrous polyethylene terephthalate and polybutylene terephthalate An outer layer; (ii) a non-nanofibrillar membrane positioned between an inner layer and the outer layer, the membrane comprising: the non-nanofibrillar membrane comprising nanopores sized to prevent passage of cells across the membrane. A membrane and (iii) Nanofibrous polyethylene terephthalate and polybutylene terephthalate and an inner layer comprising:

39. The device according to any one of claims 28 to 38, wherein the membrane pores have a diameter of 1 μm or less. Vice.

40. 40. The membrane according to any one of claims 28 to 39, wherein the membrane pores have a diameter of 0.5 μm or less. device.

41. 40. Any one of claims 28 to 39, wherein the membrane pores have a diameter of about 0.2 to 0.6 μm. The device described in

42. 40. The membrane of claim 28, wherein the pores have a diameter of about 0.4 μm. device.

43. The cell chamber further includes a loading port for loading cells into the cell chamber. Item 10. The device according to any one of items 1 to 4.

44. 10. The device of claim 1, wherein the device comprises a total thickness of 250 μm or less. device.

45. 10. The device of claim 1, wherein the device comprises a total thickness of 150 μm or less. device.

46. The cell chamber contains up to 1 x 10 ９ The size of the cell is large enough to accommodate 10. The device of claim 9.

47. The cell chamber contains up to 1 x 10 ７ The size of the cell is large enough to accommodate 10. The device of claim 9.

48. 1. A device comprising a multi-layer scaffold surrounding a cell chamber, said multi-layer scaffold comprising nanofibers an outer layer comprising resistant polyethylene terephthalate and polybutylene terephthalate; and and an inner layer comprising a soluble polyurethane, wherein the outer layer and the inner layer have a diameter of 1 μm or less. The device further comprises a nanopore.

49. 10. The device of claim 1, wherein the cell chamber contains cells.

50. 50. The device of claim 49, wherein the cells adhere to the inner layer of the scaffold.

51. 51. The device of claim 49 or 50, wherein the cells comprise retinal pigment epithelial cells.

52. The device of claim 51, wherein the cells comprise ARPE-19 cells.

53. The device is about 1×10 ５ cells to approximately 1 x 10 ８ 49 to 50 cells 52. A device according to any one of claims 52.

54. The device is about 1×10 ６ cells to approximately 1 x 10 ７ 49 to 50 cells 53. A device according to any one of claims 53.

55. 55. The method of claim 49, wherein the cells secrete a recombinant peptide or protein.

1. The device according to claim 1.

56. The cells may contain an antibody, or an antigen-binding portion thereof, a growth factor, a hormone, a prostaglandin, from the group consisting of: a medicament, an enzyme, a cytokine, a peptide therapeutic, or a combination thereof 56. The device of claim 55, which secretes a selected protein.

57. 57. The device of claim 55 or 56, wherein the cells secrete a peptide therapeutic.

58. 57. The device of claim 55 or 56, wherein the cells secrete an enzyme.

59. 59. The device of claim 58, wherein the enzyme is laronidase.

60. 59. The device of claim 58, wherein the enzyme is idursulfase.

61. 59. The device of claim 58, wherein the enzyme is arylsulfatase A.

62. 57. The method of claim 55 or 56, wherein the cell secretes an antibody, or an antigen-binding portion thereof. device.

63. 63. The method of claim 62, wherein the cell secretes a chimeric antibody, or an antigen-binding portion thereof. Vice.

64. 63. The method of claim 62, wherein the cell secretes a humanized antibody, or an antigen-binding portion thereof. Vice.

65. 63. The device of claim 62, wherein the cell secretes a human antibody, or an antigen-binding portion thereof. Chair.

66. 66. The method of claim 62, wherein the cells secrete a monoclonal antibody. device.

67. The cells contain Fab, F(ab') ２ , scFv, tandem scFv, diabody, and secreting antibody fragments selected from the group consisting of minibodies and single domain antibodies. Item 66. A device according to any one of items 62 to 65.

68. The cells express α4β7, integrin β7, TNFα, IL-12, IL-23, or or an antibody that specifically binds to an antigen selected from the group consisting of CD20, 68. The device of any one of claims 62 to 67, which secretes the moiety.

69. The cells are capable of expressing vedolizumab, abrilumab, adalimumab, Trolizumab, certolizumab ), golimumab, ustekinumab, infliximab, rituximab, and natalizumab 69. The device of claim 68, which secretes an antibody selected from:

70. the cells secrete an antibody, or an antigen-binding portion thereof, that specifically binds to α4β7. A device according to any one of claims 62 to 67.

71. 71. The method of claim 70, wherein the cell secretes vedolizumab, or an antigen-binding portion thereof. device.

72. 51. The method of claim 49 or 50, wherein the cell chamber contains cells having a three-dimensional structure. device.

73. 73. The device of claim 72, wherein the cells having a three-dimensional structure comprise a tissue explant.

74. 74. The device of claim 73, wherein the tissue is liver tissue or pancreatic tissue.

75. 73. The method of claim 72, wherein the cells having a three-dimensional structure comprise an organoid or a spheroid. The device described in

76. 76. The method of claim 75, wherein the organoid comprises hepatocytes, liver cells, or pancreatic cells. device.

77. 77. The method of claim 75 or 76, wherein the organoids are organized around sinusoids or ducts. Devices listed.

78. 10. A method for delivering a biomolecule to a subject, comprising administering to said subject a biomolecule ... and administering the device described above, wherein the cell chamber of the device contains the biomolecule. The method further comprises a cell that secretes the

79. 1. A method for delivering a recombinant peptide or protein to a subject, comprising administering to said subject:

62. The method comprising administering a device according to any one of claims 55 to 61.

80. 9. A method for delivering an antibody, or antigen-binding portion thereof, to a subject, comprising administering to said subject 72. The method comprising administering a device according to any one of claims 2 to 71.

81. The device may be implanted under the skin (subcutaneous implantation), on the omentum, in the liver, in the brain, or in the spinal canal.

81. The method of any one of claims 78 to 80, wherein the method is administered to the subject by implantation at a site selected from the group consisting of:

1. The method according to claim 1.

82. 82. The method of claim 78, wherein the device is implanted in the subject for at least 30 days.

10. The method according to any one of claims 1 to 9.

83. 82. The method of claim 78, wherein the device is implanted in the subject for at least 90 days.

10. The method according to any one of claims 1 to 9.

84. Claims 78-81, wherein the device is implanted in the subject for at least 120 days.

10. The method according to any one of claims 1 to 9.

85. 82. The method of claim 78, wherein the device is implanted in the subject for at least one year. The method according to any one of claims 1 to 4.

86. The plasma concentration of the biomolecule, recombinant peptide, or recombinant protein is determined in the subject.

78. The method of claim 77, wherein the concentration of the hydroxybenzoates in the hydroxybenzoates is at least 5 μg / mL for at least 60 days after implantation. 79, and the method according to any one of 81 to 85.

87. the plasma concentration of the antibody, or antigen-binding portion thereof, is increased in the subject at least once a day after implantation.

86. The method according to any one of claims 78 to 85, wherein the concentration is at least 5 μg / mL for at least 60 days. The method described.

88. 1. A method of treating a subject having Crohn's disease or ulcerative colitis, comprising administering to the subject a The method comprises administering the device of claim 70 or 71.

89. 70. A method of treating a subject with primary sclerosing cholangitis, comprising administering to the subject a compound according to claim 70 or 71. or 71.

90. 70. A method of treating a subject with eosinophilic esophagitis, comprising administering to the subject a compound according to claim 70 or 72. The method comprising administering the device of claim 71.

91. 70. A method of treating a subject with autoimmune hepatitis, comprising administering to the subject a compound according to claim 70 or 72. The method comprising administering the device of claim 71.

92. 58. A method of treating a subject with short bowel syndrome, comprising administering to the subject the drug of claim 57. The method comprises administering a drug.

93. A method of treating a subject with mucopolysaccharidosis type I (MPS I), comprising administering to the subject a The method comprising administering the device of claim 59.

94. A method of treating a subject with mucopolysaccharidosis type II (MPS II), comprising administering to said subject 61. The method comprising administering the device of claim 60.

95. 1. A method of treating a subject having metachromatic leukodystrophy (MLD), comprising administering to the subject 62. The method of claim 61, comprising administering the device of claim 61 to

96. A device comprising a single layer polymer scaffold surrounding a cell chamber, said scaffold comprising nano The device comprises a fibrous polymer.

97. The scaffold is made of nanofibrous polyethylene terephthalate and polybutylene terephthalate.

97. The device of claim 96, comprising:

98. 97. The device of claim 96, wherein the scaffold comprises nanofibrous polyurethane.

99. 99. The device of any one of claims 96 to 98, wherein the scaffold further comprises an anti-inflammatory agent. 。

100. The anti-inflammatory agent is tacrolimus, pirfenidone, or roflumilast. Item 99. The device of item 99.

101. 101. The device of any one of claims 96 to 100, wherein the device further comprises a population of cells. device.

102. 102. The device of claim 101, wherein the population of cells is organized in a three-dimensional structure.

103. 103. The device of claim 102, wherein the population of cells comprises a tissue explant.

104. 104. The device of claim 103, wherein the tissue explant is derived from liver tissue or pancreatic tissue. vinegar.

105. The device of claim 102, wherein the population of cells comprises an organoid.

106. 106. The method of claim 105, wherein the organoid comprises hepatocytes, liver cells, or pancreatic cells. device.

107. 105 or 1, wherein the organoids are organized around sinusoids or ducts.

06. The device according to claim 06.

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