IMPLANTABLE CELL CHAMBER DEVICE AND USES THEREOF - Patent application

Abstract

The present invention relates to an implantable cell chamber device that can retain cells but allow diffusion of biomolecules. Also disclosed is a method for continuous or semi-continuous delivery of therapeutic biomolecules to a subject in need thereof by administering to the subject a cell chamber device containing cells that secrete the therapeutic biomolecules.

Description

[Technical Field]

[0001] Related Applications This application claims priority to 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.

[0002] Sequence Listing This application contains a Sequence Listing that has been submitted electronically in ASCII format and is incorporated by reference herein in its entirety. The ASCII copy was created on March 11, 2021, is named T103022_1130WO_0465_1_SL.txt, and is 66,553 bytes in size.

[0003] The present invention relates to an implantable cell chamber device and related uses for delivering therapeutic biomolecules to subjects in need thereof. [Background technology]

[0004] Advances in biotechnology have made it possible to produce a variety of therapeutic biomolecules, such as proteins, for pharmaceutical use using recombinant DNA technology. Biomolecules are typically delivered by subcutaneous injection or intravenous infusion at regular intervals. The bioavailability of a molecule between each bolus dose is a function of the rates of absorption, distribution, and elimination; bioavailability typically peaks shortly after administration and then tapers off. The pharmacokinetic (PK) profile of each biomolecule generally determines the interval between doses, with the goal of maintaining plasma drug concentrations within a therapeutic window between toxicity and minimal efficacy. The need for frequent administration can be costly and inconvenient for patients. Additionally, the PK profile and administration frequency of each biomolecule can cause drug concentrations in each patient to fluctuate between very high and very low concentrations, depending on the amount of biomolecule present in each dose and the time interval between doses.

[0005] Because biomolecules can be larger and more complex than traditional inorganic drugs (i.e., possessing multiple functional groups in addition to complex three-dimensional structures), formulating such biomolecules can pose special challenges. For example, for a protein to maintain biological activity, the formulation must preserve the conformational integrity of at least one core sequence of amino acids in the protein while simultaneously protecting the protein's multiple functional groups from degradation. Proteins can suffer from a lack of stability during storage, and monoclonal and polyclonal antibodies, in particular, can be relatively unstable (e.g., Wang, et al., J. Pharm Sci. 96:1-26 (2007)). Consequences of chemical or physical instability of therapeutic proteins include reduced effective doses, decreased safety of therapy due to, for example, irritation or immune reactivity, and more frequent manufacturing due to a short shelf life. Numerous formulation options are available, and no single approach or system is suitable for all proteins.

[0006] Thus, there is a need to find suitable means for delivering therapeutic biomolecules to patients that will result in stable, therapeutically effective blood levels of the biomolecule over an extended period of time, in a stable and convenient form. Summary of the Invention

[0007] Provided herein is a cell chamber device capable of retaining cells while simultaneously allowing the exchange of biomolecules between the interior and exterior of the device. The device can be used to culture cells that perform functions, including, for example, the secretion of therapeutic biomolecules. When implanted in a subject, the cell chamber device can provide a stable level of biomolecule delivery to the subject over an extended period of time. Additionally or alternatively, cells within the chamber can perform physiological functions, such as removing toxins or performing metabolic operations, through the exchange of components between tissue surrounding or adjacent to the device and the cells growing within the device. The chamber includes a layered scaffold composed of a material that induces little or no fibrotic response in a mammalian host, induces little or no host immune response, retains cells inside the chamber without cell leakage into surrounding host tissue, prevents host cells from infiltrating into the chamber, and / or allows the delivery of therapeutic doses of biomolecules to the host, e.g., into the bloodstream.

[0008] Thus, in one aspect, provided herein is a device comprising a multilayer scaffold surrounding a cell chamber, the multilayer 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. In some embodiments, the nanofibrous polymer can comprise, for example, a nanofibrous polyester.

[0009] In some embodiments, the outer layer comprises nanofibrous polyethylene terephthalate or nanofibrous polybutylene terephthalate. In some embodiments, the outer layer comprises a blend of nanofibrous polyethylene terephthalate and polybutylene terephthalate. In certain embodiments, the outer layer comprises electrospun polyethylene terephthalate, electrospun polybutylene terephthalate, or a blend of electrospun polyethylene terephthalate and polybutylene terephthalate. Other suitable polymers are described herein, including, for example, nanofibrous polymethylene terephthalate or nanofibrous polyurethane.

[0010] In some embodiments, the inner layer comprises a nanofibrous polyurethane. In certain embodiments, the inner layer comprises an electrospun polyurethane.

[0011] In some embodiments, the inner layer comprises nanofibrous polyethylene terephthalate, nanofibrous polybutylene terephthalate, or a blend of nanofibrous polyethylene terephthalate and polybutylene terephthalate.

[0012] In some embodiments, the inner layer comprises electrospun polyethylene terephthalate, electrospun polybutylene terephthalate, or a blend of 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 certain embodiments, the scaffold is treated with ethylenediamine.

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

[0016] In some embodiments, the outer layer and / or inner layer comprises an anti-inflammatory agent. In some embodiments, the anti-inflammatory agent is a calcineurin inhibitor. In one embodiment, the anti-inflammatory agent is tacrolimus. In some embodiments, the anti-inflammatory agent is a pyridone. In one embodiment, the anti-inflammatory agent is pirfenidone. 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 comprises a nanopore. In certain embodiments, the nanopore is sized to allow the passage of a biomolecule. In certain embodiments, the biomolecule is 250 kilodaltons (kDa) or less.

[0018] In some embodiments, the nanopores are sized to allow passage of antibodies, or antigen-binding portions thereof. In some embodiments, the nanopores are sized to prevent passage of cells. In some embodiments, the nanopores are sized to prevent cells on one side of the multilayer scaffold from contacting cells on the other side of the multilayer scaffold.

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

[0020] In some embodiments, the multilayer scaffold further comprises a nanoporous membrane positioned between the inner and outer layers. In some embodiments, the nanoporous membrane is a solid, i.e., non-nanofiberous, polymer layer. In other embodiments, the nanoporous membrane can comprise an additional layer of nanofibrous polymer. In some embodiments, the nanoporous membrane comprises polybutylene terephthalate. In some embodiments, the nanoporous membrane comprises a layer of solid, i.e., non-nanofiberous, polybutylene terephthalate. In certain embodiments, the nanoporous membrane comprises nanofibrous polybutylene terephthalate. In some embodiments, the nanoporous membrane comprises membrane nanopores sized to allow passage of biomolecules. In certain embodiments, the biomolecules are 250 kDa or less.

[0021] In some embodiments, the membrane pores are sized to allow passage of antibodies, or antigen-binding portions thereof. In some embodiments, the membrane pores are sized to prevent passage of cells. In some embodiments, the membrane pores are sized to prevent cells on one side of the multilayer scaffold from contacting cells on the other side of the multilayer scaffold.

[0022] In another aspect, provided herein is a device comprising a multilayer scaffold surrounding a cell chamber, the multilayer scaffold comprising: (i) an outer layer comprising nanofibrous polyethylene terephthalate and polybutylene terephthalate; (ii) a non-nanofiberous membrane positioned between the inner and outer layers, the membrane comprising submicron and / or nanopores sized to prevent passage of cells across the membrane; and (iii) an inner layer comprising nanofibrous polyurethane.

[0023] In a further aspect, provided herein is a device comprising a multilayer scaffold surrounding a cell chamber, the multilayer scaffold comprising: (i) an outer layer comprising nanofibrous polyethylene terephthalate and polybutylene terephthalate; (ii) a non-nanofiberous membrane positioned between the inner and outer layers, the membrane comprising nanopores sized to prevent passage of cells across the membrane; and (iii) an inner layer comprising nanofibrous polyethylene terephthalate and polybutylene terephthalate.

[0024] In some embodiments, the membrane nanopore has a diameter of 1 μm or less. In some embodiments, the membrane nanopore has a diameter of 0.5 μm or less. In some embodiments, the membrane pore has a diameter of about 0.2-0.6 μm. In some embodiments, the membrane pore has a diameter of about 0.4 μm.

[0025] In some embodiments, the device further comprises a loading port that allows cells to be loaded into the cell chamber.

[0026] In some embodiments, the device comprises a total thickness of 250 μm or less, hi some embodiments, 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 In some embodiments, the cell chamber is sized to accommodate up to 1 x 10 cells. 7 It is sized to accommodate cells.

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

[0029] In some embodiments, the device comprises a polymer film having a pore size of 1 μm or less positioned between an outer layer and an inner layer of the scaffold.

[0030] In some embodiments, the device comprises a polymer film having a pore size of 1 μm or less, one side of the film coated with a first electrospun polymer and the other side of the film coated with a second electrospun polymer. In some embodiments, the first electrospun polymer and the second electrospun polymer comprise the same polymer. In other embodiments, the first electrospun polymer and the second electrospun polymer comprise different polymers. In some embodiments, the first electrospun polymer and / or the second electrospun polymer comprise nPET, nPBT, or a combination thereof. In some embodiments, the first electrospun polymer and / or the second electrospun polymer comprise nPU. In some embodiments, the first electrospun polymer comprises nPET-PBT and the second electrospun polymer comprises nPU.

[0031] In some embodiments, the polymer film comprises poly(ethylene terephthalate). In embodiments, the polymer film comprises pores having a diameter of about 0.2 μm to 1 μm. In various embodiments, the polymer film comprises pores having a diameter of about 0.2 μm, 0.3 μm, 0.4 μm, 0.5 μm, 0.6 μm, 0.7 μm, 0.8 μm, 0.9 μm, or 1.0 μm. In some embodiments, the polymer film comprises pores having a diameter of about 0.4 μm.

[0032] In some embodiments, the cell chamber comprises cells. In some embodiments, the cells adhere to the inner layer of the scaffold. In some embodiments, the cells comprise retinal pigment epithelial cells. In certain embodiments, the cells comprise ARPE-19 cells. In some embodiments, the cells comprise hepatocytes. 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 cells.

[0034] In some embodiments, the device has a capacitance of about 1×10 6 cells ~ approx. 1 x 10 7 Contains cells.

[0035] In some embodiments, the cells secrete a recombinant peptide or protein, hi certain embodiments, the cells secrete a protein selected from the group consisting of a growth factor, a hormone, a cytokine, a prostaglandin, an enzyme, or a combination thereof.

[0036] In some embodiments, the cells secrete a peptide therapeutic (eg, a peptide therapeutic for gastrointestinal use, such as treating short bowel syndrome).

[0037] In some embodiments, the cells secrete an enzyme. In one embodiment, the enzyme is idursulfase. 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 certain embodiments, the cells secrete chimeric antibodies, or antigen-binding portions thereof. In some embodiments, the cells secrete humanized antibodies, or antigen-binding portions thereof. In some embodiments, the cells secrete human antibodies, or antigen-binding portions thereof. In some embodiments, the cells secrete monoclonal antibodies. In some embodiments, the cells secrete antibody fragments selected from the group consisting of Fab, F(ab')2, scFv, tandem scFv, diabodies, minibodies, and single domain antibodies.

[0039] In some embodiments, the cells secrete an antibody, or an antigen-binding portion thereof, that specifically binds to an antigen selected from the group consisting of α4β7 integrin, integrin β7, TNFα, IL-12, IL-23, or CD20. In some embodiments, the cells secrete an antibody selected from vedolizumab, abrilumab, adalimumab, etrolizumab, certolizumab, golimumab, ustekinumab, infliximab, rituximab, and natalizumab.

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

[0041] In some embodiments, the cell chamber comprises a population of cells having a three-dimensional structure. In one embodiment, the population of cells having a three-dimensional structure is a tissue explant. In some embodiments, the tissue is liver tissue, kidney tissue, or pancreatic tissue. In other embodiments, the population of cells having a three-dimensional structure is organized as a spheroid. In some embodiments, the spheroid comprises hepatocytes, liver cells, or pancreatic cells. In certain embodiments, the spheroid is organized around a sinusoid or duct.

[0042] In another aspect, provided herein is a method of delivering a biomolecule to a subject, the method comprising administering to the subject a device as described herein, wherein the cell chamber of the device contains cells that secrete the biomolecule.

[0043] In another aspect, provided herein is a method of delivering a recombinant peptide or protein to a subject, the method comprising administering to the subject a device of the present disclosure.

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

[0045] In some embodiments, the device is administered to the subject by implantation at a site selected from under the skin (subcutaneous implantation), on the omentum, in the liver, in the brain, or in 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. In some embodiments, 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 5 μg / mL in the subject for a period of at least 60 days after implantation.

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

[0051] In another aspect, provided herein is a method of treating a subject with primary sclerosing cholangitis, the method comprising administering to the subject a device of the present disclosure.

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

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

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

[0055] In another aspect, provided herein is a method of treating a subject with Mucopolysaccharidosis Type I (MPS I), comprising administering to the subject a device of the present disclosure (e.g., a device in which cells secrete an enzyme such as laronidase). In certain embodiments, the device comprises cells that secrete an enzyme comprising the amino acid sequence of SEQ ID NO:50, or an enzyme having at least 90%, 92%, 94%, 95%, 96%, 98%, or 99% identity to SEQ ID NO:50.

[0056] In another aspect, provided herein is a method of treating a subject with Mucopolysaccharidosis Type II (MPS II), comprising administering to the subject a device of the present disclosure (e.g., a device in which cells secrete an enzyme such as idursulfase). In certain embodiments, the device comprises cells that secrete an enzyme comprising the amino acid sequence of SEQ ID NO:51, or an enzyme having at least 90%, 92%, 94%, 95%, 96%, 98%, or 99% identity to SEQ ID NO:51.

[0057] In another aspect, provided herein is a method of treating a subject with metachromatic leukodystrophy (MLD), comprising administering to the subject a device of the present disclosure (e.g., a device in which cells secrete an enzyme such as arylsulfatase A). In certain embodiments, the device comprises cells that secrete an enzyme comprising the amino acid sequence of SEQ ID NO:52, or an enzyme having at least 90%, 92%, 94%, 95%, 96%, 98%, or 99% identity to SEQ ID NO:52.

[0058] In another aspect, provided herein is a device comprising a monolayer scaffold surrounding a cell chamber. Such cell chamber devices, in some embodiments, can be used to culture cells growing in three-dimensional culture or as tissue explants. In some embodiments, the cell chamber can contain cells having a three-dimensional structure. In some embodiments, the monolayer scaffold can comprise a nanofibrous polymer as described herein. In some embodiments, the nanofibrous polymer is a polyester, such as nanofibrous polyethylene terephthalate or nanofibrous polybutylene terephthalate. In some embodiments, the monolayer scaffold comprises a blend of nanofibrous polyethylene terephthalate and polybutylene terephthalate. In some embodiments, the monolayer scaffold comprises a nanofibrous polyurethane.

[0059] In some embodiments, the cell chamber contains cells having a three-dimensional structure. In one embodiment, the cells having a three-dimensional structure are tissue explants. In some embodiments, the tissue is liver tissue, kidney tissue, or pancreatic tissue. In other embodiments, the cells having a three-dimensional structure are organized as spheroids. In some embodiments, the spheroids can comprise hepatocytes, liver cells, or pancreatic cells. In certain embodiments, the spheroids are organized around sinusoids or ducts. In some embodiments, the cells are hepatocytes. In other embodiments, the cells are pancreatic islet cells. It should be noted that the present invention may include the following aspects. [Aspect 1] A device comprising a multi-layered scaffold surrounding a cell chamber, the multi-layered 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. [Aspect 2] 10. The device of embodiment 1, wherein the outer layer comprises nanofibrous polyethylene terephthalate and polybutylene terephthalate. [Aspect 3] 10. The device of embodiment 1, wherein the outer layer comprises electrospun polyethylene terephthalate and polybutylene terephthalate. [Aspect 4] Aspect 4. The device of any one of aspects 1-3, wherein the inner layer comprises nanofibrous polyurethane. [Aspect 5] 5. The device of embodiment 4, wherein the inner layer comprises electrospun polyurethane. [Aspect 6] Aspect 4. The device of any one of aspects 1-3, wherein the inner layer comprises nanofibrous polyethylene terephthalate and polybutylene terephthalate. [Aspect 7] Aspect 4. The device of any one of aspects 1-3, wherein the inner layer comprises electrospun polyethylene terephthalate and polybutylene terephthalate. [Aspect 8]

[0023] Aspect 1, the device of any one of the preceding aspects, wherein the outer layer and / or the inner layer comprises one or more charged surface modifications. [Aspect 9] 9. The device of embodiment 8, wherein the outer layer and / or the inner layer have a net positive charge. [Aspect 10] 10. The device of embodiment 9, wherein the scaffold is treated with ethylenediamine. [Aspect 11] 9. The device of embodiment 8, wherein the outer layer and / or the inner layer have a net negative charge. [Aspect 12] 12. The device of embodiment 11, wherein the scaffold has been treated with sodium hydroxide. [Aspect 13] 20. The device of any one of the preceding aspects, wherein the outer layer and / or the inner layer comprises an anti-inflammatory agent. [Aspect 14] Aspect 14. The device of aspect 13, wherein the anti-inflammatory agent is a calcineurin inhibitor. [Aspect 15] 15. The device of aspect 14, wherein the anti-inflammatory agent is tacrolimus. [Aspect 16] 14. The device of embodiment 13, wherein the anti-inflammatory agent is a pyridone. [Aspect 17] 17. The device of embodiment 16, wherein the anti-inflammatory agent is pirfenidone. [Aspect 18] Aspect 14. The device of aspect 13, wherein the anti-inflammatory agent is a phosphodiesterase inhibitor. [Aspect 19] 20. The device of embodiment 18, wherein the anti-inflammatory agent is roflumilast. [Aspect 20]

[0023] Aspect 12. The device of any one of the preceding aspects, wherein the outer layer and / or the inner layer comprise pores. [Aspect 21] 21. The device of embodiment 20, wherein the pores are sized to allow the passage of biomolecules. [Aspect 22] 22. The device of embodiment 21, wherein the biomolecule is 250 kDa or less. [Aspect 23] 21. The device of embodiment 20, wherein the pores are sized to allow the passage of an antibody, or antigen-binding portion thereof. [Aspect 24] 24. The device of any one of aspects 20 to 23, wherein the pores are sized to prevent the passage of cells. [Aspect 25] 25. The device of any one of aspects 20-24, wherein the pores are sized to prevent cells on one side of the multilayer scaffold from contacting cells on the other side of the multilayer scaffold. [Aspect 26] 26. The device of any one of embodiments 20 to 25, wherein the pores have a diameter of 1 μm or less. [Aspect 27] 27. The device of any one of embodiments 20 to 26, wherein the pores have a diameter of 0.5 μm or less. [Aspect 28]

[0023] Aspect 12. The device of any one of the preceding aspects, wherein the multilayered scaffold further comprises a porous membrane positioned between the inner layer and the outer layer. [Aspect 29] 29. The device of embodiment 28, wherein the porous membrane comprises polyethylene terephthalate. [Aspect 30] 30. The device of embodiment 29, wherein the porous membrane comprises nanofibrous polyethylene terephthalate. [Aspect 31] 30. The device of embodiment 29, wherein the nanoporous membrane comprises non-nanofiberous polyethylene terephthalate. [Aspect 32] 32. The device of any one of aspects 28 to 31, wherein the porous membrane comprises membrane pores sized to allow passage of the biomolecules. [Aspect 33] 33. The device of embodiment 32, wherein the biomolecule is 250 kDa or less. [Aspect 34] Aspect 34. The device of any one of aspects 28 to 33, wherein the membrane pores are sized to allow the passage of the antibody, or antigen-binding portion thereof. [Aspect 35] Aspect 35. The device of any one of aspects 28 to 34, wherein the membrane pores are sized to prevent the passage of the cells. [Aspect 36] 36. The device of any one of aspects 28 to 35, wherein the membrane pores are sized to prevent cells on one side of the multilayer scaffold from contacting cells on the other side of the multilayer scaffold. [Aspect 37] 1. A device comprising a multi-layer scaffold surrounding a cell chamber, said multi-layer scaffold comprising: (i) an outer layer comprising nanofibrous polyethylene terephthalate and polybutylene terephthalate; (ii) a non-nanofibrillar membrane positioned between an inner layer and the outer layer, the membrane comprising nanopores sized to prevent passage of cells across the membrane; (iii) an inner layer comprising nanofibrous polyurethane. [Aspect 38] 1. A device comprising a multi-layer scaffold surrounding a cell chamber, said multi-layer scaffold comprising: (i) an outer layer comprising nanofibrous polyethylene terephthalate and polybutylene terephthalate; (ii) a non-nanofibrillar membrane positioned between an inner layer and the outer layer, the membrane comprising nanopores sized to prevent passage of cells across the membrane; (iii) an inner layer comprising nanofibrous polyethylene terephthalate and polybutylene terephthalate. [Aspect 39] Aspect 39. The device of any one of aspects 28 to 38, wherein the membrane pores have a diameter of 1 μm or less. [Aspect 40] Aspect 40. The device of any one of aspects 28 to 39, wherein the membrane pores have a diameter of 0.5 μm or less. [Aspect 41] 40. The device of any one of embodiments 28 to 39, wherein the membrane pores have a diameter of about 0.2 to 0.6 μm. [Aspect 42] 40. The device of any one of embodiments 28 to 39, wherein the membrane pores have a diameter of about 0.4 μm. [Aspect 43]

[0023] Aspect 12. The device of any one of the preceding aspects, further comprising a loading port that allows cells to be loaded into the cell chamber. [Aspect 44] The device of any one of the preceding aspects, wherein the device comprises a total thickness of 250 μm or less. [Aspect 45] The device of any one of the preceding aspects, wherein the device comprises a total thickness of 150 μm or less. [Aspect 46] The cell chamber contains up to 1 x 10 9

[0023] The device of any one of the preceding aspects, wherein the device is sized to accommodate cells. [Aspect 47] The cell chamber contains up to 1 x 10 7

[0023] The device of any one of the preceding aspects, wherein the device is sized to accommodate cells. [Aspect 48] A device comprising a multi-layered scaffold surrounding a cell chamber, the multi-layered scaffold comprising an outer layer comprising nanofibrous polyethylene terephthalate and polybutylene terephthalate and an inner layer comprising nanofibrous polyurethane, the outer layer and the inner layer comprising nanopores having a diameter of 1 μm or less. [Aspect 49]

[0023] Aspect 15. The device of any one of the preceding aspects, wherein the cell chamber contains cells. [Aspect 50] 50. The device of embodiment 49, wherein the cells adhere to the inner layer of the scaffold. [Aspect 51] 51. The device of embodiment 49 or 50, wherein the cells comprise retinal pigment epithelial cells. [Aspect 52] 52. The device of embodiment 51, wherein the cells comprise ARPE-19 cells. [Aspect 53] The device is about 1×10 5 cells ~ approx. 1 x 10 8 53. The device of any one of embodiments 49 to 52, comprising cells. [Aspect 54] The device is about 1×10 6 cells ~ approx. 1 x 10 7 54. The device of any one of embodiments 49 to 53, comprising cells. [Aspect 55] 55. The device of any one of aspects 49 to 54, wherein the cells secrete a recombinant peptide or protein. [Aspect 56] 56. The device of embodiment 55, wherein the cells secrete a protein selected from the group consisting of an antibody, or antigen-binding portion thereof, a growth factor, a hormone, a prostaglandin, an enzyme, a cytokine, a peptide therapeutic, or a combination thereof. [Aspect 57] 57. The device of embodiment 55 or 56, wherein the cells secrete a peptide therapeutic. [Aspect 58] 57. The device of embodiment 55 or 56, wherein the cells secrete an enzyme. [Aspect 59] 59. The device of embodiment 58, wherein the enzyme is laronidase. [Aspect 60] 59. The device of embodiment 58, wherein the enzyme is idursulfase. [Aspect 61] 59. The device of embodiment 58, wherein the enzyme is arylsulfatase A. [Aspect 62] 57. The device of embodiment 55 or 56, wherein the cells secrete antibodies, or antigen-binding portions thereof. [Aspect 63] 63. The device of embodiment 62, wherein the cells secrete a chimeric antibody, or an antigen-binding portion thereof. [Aspect 64] 63. The device of embodiment 62, wherein the cells secrete a humanized antibody, or an antigen-binding portion thereof. [Aspect 65] 63. The device of embodiment 62, wherein the cells secrete human antibodies, or antigen-binding portions thereof. [Aspect 66] 66. The device of any one of aspects 62 to 65, wherein the cells secrete a monoclonal antibody. [Aspect 67] The cells are Fab, F(ab') 2 66. The device of any one of aspects 62 to 65, wherein the device secretes an antibody fragment selected from the group consisting of an scFv, a tandem scFv, a diabody, a minibody, and a single domain antibody. [Aspect 68] 68. The device of any one of aspects 62-67, wherein the cells secrete an antibody, or an antigen-binding portion thereof, that specifically binds to an antigen selected from the group consisting of α4β7, integrin β7, TNFα, IL-12, IL-23, or CD20. [Aspect 69] 70. The device of embodiment 68, wherein the cells secrete an antibody selected from vedolizumab, abrilumab, adalimumab, etrolizumab, certolizumab, golimumab, ustekinumab, infliximab, rituximab, and natalizumab. [Aspect 70] 68. The device of any one of aspects 62 to 67, wherein the cells secrete an antibody, or an antigen-binding portion thereof, that specifically binds to α4β7. [Aspect 71] 71. The device of embodiment 70, wherein the cells secrete vedolizumab, or an antigen-binding portion thereof. [Aspect 72] 51. The device of embodiment 49 or 50, wherein the cell chamber contains cells having a three-dimensional structure. [Aspect 73] 73. The device of embodiment 72, wherein the cells having a three-dimensional structure comprise a tissue explant. [Aspect 74] The device of embodiment 73, wherein the tissue is liver tissue or pancreatic tissue. [Aspect 75] 73. The device of embodiment 72, wherein the cells having a three-dimensional structure comprise organoids or spheroids. [Aspect 76] 76. The device of embodiment 75, wherein said organoids comprise hepatocytes, liver cells, or pancreatic cells. [Aspect 77] 77. The device of embodiment 75 or 76, wherein said organoids are organized around sinusoids or ducts. [Aspect 78] 10. A method of delivering a biomolecule to a subject, comprising administering to the subject the device of any one of the preceding aspects, wherein the cell chamber of the device contains cells that secrete the biomolecule. [Aspect 79] 62. A method of delivering a recombinant peptide or protein to a subject, the method comprising administering to the subject a device according to any one of aspects 55 to 61. [Aspect 80] 72. A method of delivering an antibody, or an antigen-binding portion thereof, to a subject, comprising administering to the subject a device according to any one of aspects 62 to 71. [Aspect 81] 81. The method of any one of aspects 78-80, wherein the device is administered to the subject by implantation at a site selected from under the skin (subcutaneous implantation), on the omentum, in the liver, in the brain, or in the spinal cord canal. [Aspect 82] 82. The method of any one of aspects 78-81, wherein the device is implanted in the subject for at least 30 days. [Aspect 83] 82. The method of any one of aspects 78-81, wherein the device is implanted in the subject for at least 90 days. [Aspect 84] 82. The method of any one of aspects 78-81, wherein the device is implanted in the subject for at least 120 days. [Aspect 85] 82. The method of any one of aspects 78-81, wherein the device is implanted in the subject for at least one year. [Aspect 86] 86. The method of any one of aspects 78, 79, and 81-85, wherein the plasma concentration of the biomolecule, recombinant peptide, or recombinant protein is at least 5 μg / mL in the subject for at least 60 days after implantation. [Aspect 87] 86. The method of any one of aspects 78 to 85, wherein the plasma concentration of the antibody, or antigen-binding portion thereof, is at least 5 μg / mL in the subject for at least 60 days post-implantation. [Aspect 88] 72. A method of treating a subject having Crohn's disease or ulcerative colitis, comprising administering to the subject a device of aspect 70 or 71. [Aspect 89] 72. A method of treating a subject having primary sclerosing cholangitis, comprising administering to the subject a device of aspect 70 or 71. [Aspect 90] 72. A method of treating a subject having eosinophilic esophagitis, comprising administering to the subject a device of aspect 70 or 71. [Aspect 91] 72. A method of treating a subject having autoimmune hepatitis, comprising administering to the subject a device of aspect 70 or 71. [Aspect 92] 60. A method of treating a subject having short bowel syndrome, comprising administering to the subject a device of aspect 57. [Aspect 93] 60. A method of treating a subject having Mucopolysaccharidosis Type I (MPS I), comprising administering to the subject a device of aspect 59. [Aspect 94] 61. A method of treating a subject having mucopolysaccharidosis type II (MPS II), comprising administering to the subject a device of aspect 60. [Aspect 95] 62. A method of treating a subject having metachromatic leukodystrophy (MLD), comprising administering to the subject a device according to aspect 61. [Aspect 96] A device comprising a single layer polymer scaffold surrounding a cell chamber, said scaffold comprising a nanofibrous polymer. [Aspect 97] 97. The device of embodiment 96, wherein the scaffold comprises nanofibrous polyethylene terephthalate and polybutylene terephthalate. [Aspect 98] 97. The device of embodiment 96, wherein the scaffold comprises nanofibrous polyurethane. [Aspect 99]

[0082] Aspect 99. The device of any one of aspects 96-98, wherein the scaffold further comprises an anti-inflammatory agent. [Aspect 100] Aspect 99 is a device according to aspect 99, wherein the anti-inflammatory agent is tacrolimus, pirfenidone, or roflumilast. [Aspect 101] The device of any one of aspects 96 to 100, wherein the device further comprises a population of cells. [Aspect 102] 102. The device of embodiment 101, wherein the population of cells is organized in a three-dimensional structure. [Aspect 103] 103. The device of embodiment 102, wherein the population of cells comprises a tissue explant. [Aspect 104] A device according to embodiment 103, wherein the tissue explant is derived from liver tissue or pancreatic tissue. [Aspect 105] 103. The device of embodiment 102, wherein the population of cells comprises organoids. [Aspect 106] The device of embodiment 105, wherein the organoids comprise hepatocytes, liver cells, or pancreatic cells. [Aspect 107] 107. The device of embodiment 105 or 106, wherein said organoids are organized around sinusoids or ducts. [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 a fiber having a diameter of less than 3.0 micrometers. In an exemplary embodiment, the nanofiber has a diameter of 10 nanometers to 3.0 micrometers. The diameter of the nanofiber depends on the type of polymer used and the method of production. Methods for making nanofibers include electrospinning, drawing, self-assembly, template synthesis, and thermally induced phase separation.

[0062] As used herein, the term "electrospinning" refers to a process that uses high voltage in combination with a source-to-base distance to produce a crosslinked mesh. For example, an electric field can be used to draw a solution containing a polymer from the tip of a capillary to a collector. A high-voltage DC current can be applied to the solution, thereby drawing a jet of solution toward a grounded collector screen. Upon exiting the capillary orifice, the charged solution jet can be evaporated to form fibers, which can be collected on a collector (e.g., a rotating collector). The size and morphology of the resulting fibers depend on various factors, such as the viscosity of the solution, molecular weight, the nature of the polymer, and other parameters related to the electrospinning apparatus. The electrospinning process for forming polymer nanofibers has been demonstrated using a variety of polymers (see, for example, Huang, et al., Composites Science and Technology (2003) 63:2223-2253, incorporated herein by reference in its entirety).

[0063] As used herein, "electrospun polymer" refers to a polymer fiber, e.g., a polymer nanofiber, produced by electrospinning. Electrospun polymers can include, but are not limited to, polyethylene terephthalate (PET), polybutylene terephthalate (PBT), or polyurethane (PU). The aforementioned polymers can be electrospun to produce polymer nanofibers, as described herein.

[0064] As used herein, the term "biomolecule" refers to any organic molecule that can be produced by a living organism, including polypeptides, polysaccharides, and polynucleotides, as well as organic molecules such 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 acid (DNA), or ribonucleic acid (RNA). In some embodiments, a biomolecule is recombinantly produced. In some embodiments, a biomolecule may be secreted from cells by other means, such as extracellular vesicles, exosomes, or secretory apparatus such as mitochondria. For example, secreted biomolecules may include those assembled, packaged, and secreted as exosomes, lipid polymers, or viral particles. In some embodiments, a biomolecule may be a "therapeutic biomolecule" that, when provided to a subject in an effective amount or dose, can treat, prevent, and / or ameliorate symptoms of a disease, disorder, infection, or condition in a subject, e.g., a human patient, in need thereof. In certain embodiments, cells grown in the cell chamber devices described herein can secrete an effective amount of a therapeutic biomolecule in a subject to which the device is administered. In some embodiments, the therapeutic biomolecule is an antibody. In other embodiments, the therapeutic biomolecule is a hormone. In other embodiments, the therapeutic biomolecule is an enzyme. In other embodiments, the therapeutic biomolecule is a peptide or protein.

[0065] The term "antibody," as used herein, refers to an immunoglobulin molecule composed of four polypeptide chains, two heavy (H) chains and two light (L) chains, interconnected by disulfide bonds. Each heavy chain consists of a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region (CH). The heavy chain constant region consists of three domains, CH1, CH2, and CH3. Each light chain consists of a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region consists of one domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability, called complementarity-determining regions (CDRs), interspersed with more conserved regions, called framework regions (FRs). Each VH and VL consists of three CDRs and four FRs, arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In some embodiments, the antibody has a fragment crystallizable (Fc) region. Antibodies suitable for use in the embodiments described herein may include antibodies having an isotype selected from IgG (e.g., IgG1, IgG2, IgG3, IgG4), IgM, IgA1, IgA2, IgD, or IgE. In exemplary embodiments, the antibody is an IgG1 antibody, an IgG2 antibody, or an IgG3 antibody. In various embodiments, the antibody may comprise a kappa light chain or a lambda light chain. In certain embodiments, the antibody may have an IgG1 isotype and a kappa light chain.

[0066] The cell surface molecule, "α4β7 integrin" or "α4β7" (used interchangeably throughout) is a heterodimer of the α4 chain (CD49D, ITGA4) and the β7 chain (ITGB7). Human α4-integrin and β7-integrin genes, GenBank (National Center for Biotechnology Information, Bethesda, Md.) reference sequence accession numbers NM_000885 and NM_000889, respectively, are expressed by B and T lymphocytes, particularly memory CD4+ lymphocytes. Typical of many integrins, α4β7 exists in either a resting or activated state. Ligands for α4β7 include vascular cell adhesion molecule (VCAM), fibronectin, and mucosal addressin (MAdCAM, e.g., MAdCAM-1). Antibodies that bind to α4β7 integrin are referred to herein as "anti-α4β7 antibodies."

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

[0068] "CDRs" or "complementarity determining regions" are regions of hypervariability interspersed within more conserved regions called "framework regions" (FR).

[0069] As used herein, the term "antibody fragment" or "antigen-binding fragment" of an antibody refers to Fab, Fab', F(ab')2, and Fv fragments, single-chain antibodies, functional heavy-chain antibodies (nanobodies), and any portion of an antibody that has specificity for at least one desired epitope (e.g., an isolated portion of the complementarity-determining regions with sufficient framework sequences to specifically bind to the epitope) that competes with the intact antibody for specific binding. Antigen-binding fragments can be produced by recombinant techniques or by enzymatic or chemical cleavage of antibodies. Exemplary antibody fragments include, but are not limited to, Fab, F(ab')2, scFv, tandem scFv, diabodies, minibodies, and single-domain antibodies.

[0070] "Humanized" forms of non-human (e.g., rodent) antibodies are chimeric antibodies that contain minimal sequence derived from the non-human antibody. For the most part, humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a hypervariable region of the recipient are replaced by residues from a hypervariable region of a non-human species (donor antibody) such as mouse, rat, rabbit, or non-human primate having the desired specificity, affinity, and capacity. In some instances, framework region (FR) residues of the human antibody are replaced by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues that are not found in the recipient or donor antibody. These modifications are made to further refine antibody performance. Generally, humanized antibodies will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable CDR loops correspond to those of a non-human antibody and all or substantially all of the FRs are those of a human immunoglobulin sequence. The humanized antibody optionally also will comprise at least a portion of an antibody constant region (Fc), typically that of a human antibody. For further details, see, e.g., Jones et al., Nature 321:522-525 (1986), Riechmann et al., Nature 332:323-329 (1988), and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992).

[0071] As used herein, the term "pore(s)" refers to openings in a material, e.g., a scaffolding material, that allow the passage of biomolecules from one side of the material to the other. The pores can be of any shape or dimension suitable for this purpose. When the material is a nanofibrous polymer, e.g., an electrospun nanofibrous polymer, the spaces between the fibers can function as pores. In one embodiment, the pores can be of a size that allows the passage of biomolecules but blocks the passage of cells.

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

[0073] The term "recombinant host cell" (used interchangeably herein with the term "host cell") includes cells into which a recombinant expression vector, e.g., a recombinant expression vector encoding a secreted protein, has been introduced. It is understood that such terms are intended to refer not only to the particular subject cell but also to the progeny of such a cell. Because certain modifications may occur in successive generations, either due to mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term "host cell" as used herein. Furthermore, unless otherwise specified, it is understood that when the term "cell," e.g., host cell or mammalian cell or mammalian host cell, is used, it is intended to include a population of cells.

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

[0075] II. Implantable Cell Chamber Device Provided herein is a cell chamber device capable of retaining cells, for example, cells secreting therapeutic biomolecules. The cell chamber device of the present disclosure includes a scaffold comprising a nanofibrous polymer that defines the envelope of the cell chamber. The scaffold allows the passage of biomolecules, such as proteins, into and out of the cell chamber, but not cells. Live cells capable of producing the biomolecules can be loaded into the cell chamber, where they can grow and adhere to the inner surface of the scaffold. Biomolecules secreted by cells in the cell chamber can diffuse out of the cell chamber through pores in the scaffold. The scaffold prevents the contained cells from migrating out of the cell chamber and prevents host cells from infiltrating into the cell chamber. When the cell chamber is implanted into a host subject, the cells in the cell chamber device can provide stable delivery of various biomolecular therapeutics in stable form, for example, for treatment of a host subject in need thereof. Continuous production of desired biomolecules by cells in the chamber can eliminate the need for repeated administration of isolated pharmaceutical compositions containing the biomolecules by conventional means. The cell chamber devices provided herein preferably comprise materials that induce little or no fibrotic response in a mammalian host and induce little or no host immune response.

[0076] The cell chamber devices provided herein include a scaffold comprising a nanofibrous polymer surrounding the cell chamber. In some embodiments, the scaffold of the cell chamber device has a multi-layer structure. For example, the scaffold can include two layers of nanofibrous polymer having an outer layer and an inner layer, e.g., the outer layer of nanofibrous polymer contacts the external environment surrounding the device, and the inner layer of nanofibrous polymer contacts the cell chamber. Alternatively, the scaffold can have a three-layer or multi-layer structure by including one or more additional layers between the outer and inner layers. For example, the scaffold can optionally include a membrane, e.g., a porous membrane, a semi-porous membrane, or a non-porous membrane, positioned between the outer and inner layers. The membrane is alternatively referred to herein as a filter. In some embodiments, one or more additional layers of the scaffold comprise a nanofibrous polymer. In some embodiments, the multi-layer scaffold includes 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more layers. The outer and inner layers (as well as any additional layers, e.g., the central membrane layer) may comprise the same nanofibrous polymer, or alternatively, may comprise different nanofibrous polymers with different properties. For example, the outer layer of the scaffold may comprise a material that is compatible with implantation into host tissue (e.g., elicits little or no fibrotic or immune response within the implant recipient and allows vascularization of the outer scaffold), while the inner layer, which directly contacts the cells in the cell chamber, may comprise a material that can function as a cell scaffold and optionally a barrier. In certain embodiments, the inner layer may have smaller pores than the outer layer.

[0077] The inner and / or outer scaffold layers of the cell chamber device can be formed from a variety of nanofibrous polymers, such as polyethylene terephthalate (PET, also known as Dacron), polybutylene terephthalate (PBT), or polyurethane (PU). Any biocompatible polymer suitable for use in electrospinning can be used in the cell chamber devices described herein. In some embodiments, the biocompatible polymer is a nanofibrous polyester. Other polymers suitable for use in the scaffold layer of the cell chamber devices described herein include, but are not limited to, polylactic acid (PLA), polyglycolic acid (PGA), polylactic-co-glycolic acid (PLGA), polycaprolactone (PCL), polypropylene (PP), polytetrafluoroethylene (PTFE), polytrimethylene terephthalate, polyvinyl alcohol (PVA), polyethylene oxide (PEO), polytrimethylene polyterephthalate (PTT), polyethylene acetate (PEVA), poly-D-lactide (PLDA), polylactic acid (PLLA), or polyethylene glycol (PEG). Other suitable polymers include, but are not limited to, collagen, gelatin, alginate, fibrinogen, silk, elastin, cellulose, chitin, and chitosan. The inner and / or outer layers may each comprise a single type of nanofibrous polymer. In such cases, the inner and / or outer layers may each comprise different types of nanofibrous polymers or may each comprise the same type of nanofibrous polymer. Alternatively, the inner and / or outer layers may each comprise two or more types of nanofibrous polymers.

[0078] For example, in some embodiments, the outer layer can include nanofibrous polyethylene terephthalate (nPET) or nanofibrous polybutylene terephthalate (nPBT). In some embodiments, the outer layer can include both nanofibrous polyethylene terephthalate and polybutylene terephthalate (nPET-PBT). In some embodiments, the outer layer can include nanofibrous polyurethane (nPU).

[0079] In some embodiments, the inner layer can comprise nanofibrous polyethylene terephthalate (nPET) or nanofibrous polybutylene terephthalate (nPBT). In some embodiments, the inner layer can comprise both nanofibrous polyethylene terephthalate and polybutylene terephthalate (nPET-PBT). In some embodiments, the inner layer can comprise nanofibrous polyurethane (nPU). In some embodiments, the outer layer can comprise nPET-PBT and the inner layer can comprise nPU. In other embodiments, the outer layer can comprise nPU. In some embodiments, the inner layer can comprise nPET, nPBT, or nPET-PBT.

[0080] The scaffold can additionally or alternatively comprise other synthetic or biological materials. Examples of synthetic materials include polytetrafluoroethylene (ePTFE) or poly(glycolic acid) (PGA). Biological materials include biological membranes, bovine tissue, collagen scaffolds (e.g., gels, threads, foams, sheets, mats, or tubes), gelatin, alginate, cellulose (e.g., methylcellulose), elastin, glycosaminoglycans, peptidoglycans, chitin, or fibrin (e.g., gels). The biological material can optionally be treated with a crosslinking agent, such as an aldehyde (e.g., glutaraldehyde or formaldehyde), a carbodiimide (e.g., 1-ethyl-3-(dimethylaminopropyl)carbodiimide), an acrylamide (e.g., N,N'-methylenebisacrylamide), or a diimidate (e.g., dimethylsuberimidate). In some embodiments, the biological material comprises a glutaraldehyde-crosslinked biological membrane.

[0081] There are several different methods for synthesizing tissue scaffolds that can be used to fabricate the cell chamber devices described herein. These include, for example, electrospinning, nanofiber self-assembly, textile techniques, solvent casting, and particulate leaching. One exemplary method for creating scaffold layers is electrospinning, a process in which an electric field is used to draw a solution containing a polymer through the tip of a capillary and into a collector. A high-voltage DC current is applied to the solution, which draws a jet of solution toward a grounded collector screen. Upon exiting the capillary orifice, the electrically charged solution jet evaporates, forming fibers that are collected on a collector. Electrospinning can be used to produce crosslinked meshes of polymer fibers with diameters ranging from nanometers to micrometers. The size and morphology of the fibers obtained by electrospinning depend on various factors, such as the viscosity of the solution, molecular weight, properties of the polymer, and other parameters related to the electrospinning device. The electrospinning process for forming polymer nanofibers has been demonstrated using a variety of polymers (see, e.g., Huang, et al. Composites Science and Technology (2003) 63:2223-2253, which is incorporated herein by reference in its entirety).

[0082] In certain embodiments, the inner and / or outer layers of the scaffold are produced by electrospinning, thereby forming a scaffold comprising electrospun polymer. Electrospun polymers are biocompatible and have a random web-like structure similar to the body's natural extracellular matrix scaffold, thereby promoting tissue integration and preventing rejection. Furthermore, electrospun polymers can enhance cell growth, promote cell differentiation, and attract cell attachment. Additionally, the size of electrospun fibers can be 10-14 times smaller than the fiber size of other implantable materials (e.g., woven polyester membranes, see Figures 1A-1C).

[0083] In exemplary embodiments, the scaffold (e.g., inner and / or outer layers) can comprise an electrospun polymer selected from electrospun polyethylene terephthalate, electrospun polybutylene terephthalate, or 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 comprises electrospun polyethylene terephthalate and electrospun polybutylene terephthalate (see FIG. 1C, bottom image). In further embodiments, the inner layer comprises electrospun polyurethane (see FIG. 1C, top image). In exemplary embodiments, the outer layer comprises electrospun polyethylene terephthalate and electrospun polybutylene terephthalate, and the inner layer comprises electrospun polyurethane.

[0084] Electrospun polymers can be produced by electrospanning procedures known in the art or further described herein. In one embodiment, electrospun polymers are prepared by first preparing a polymer solution, such as a polymer solution containing a polymer and hexafluoroisopropanol (HFIP). For example, to form a solution of polyethylene terephthalate (PET) and polybutylene terephthalate (PBT), PET and PBT chips or pellets can be placed in a solution of HFIP and incubated (e.g., on a rotator) until the chips dissolve (e.g., about 5-7 days of incubation). Similarly, to form a solution of polyurethane (PU), PU chips or pellets can be placed in a solution of HFIP and incubated (e.g., on a rotator) until the chips dissolve (e.g., about 5-7 days of incubation).

[0085] The polymer solution can then be loaded onto an electrospinning unit to produce an electrospun polymer. For example, a polyester solution can be loaded into a syringe, which is then connected to the electrospinning unit. The electrospinning unit can then be operated according to standard operating procedures to produce an electrospun polymer (e.g., PET, PBT, or PU). In some embodiments, the electrospinning unit is configured to operate at an electrospinning distance of about 10-25 cm (e.g., about 10 cm, about 11 cm, about 12 cm, about 13 cm, about 14 cm, about 15 cm, about 16 cm, about 17 cm, about 18 cm, about 19 cm, or about 20 cm, about 21 cm, about 22 cm, about 23 cm, about 24 cm, or about 25 cm), at about 15-30 kV (e.g., about 15 kV). The electrospun material is set to have an electrospinning voltage of about 18 kV, about 20 kV, about 21 kV, about 22 kV, or about 23 kV, about 25 kV, about 28 kV, or about 30 kV, or a rotation speed of about 15-150 RPM (e.g., about 15-25 RPM, about 25-50 RPM, about 50-75 RPM, about 75-100 RPM, about 100-125 RPM, or about 125-150 RPM). In some embodiments, a second electrospun polymer (e.g., a PU layer) can be layered on a first electrospun polymer (e.g., an nPET-PBT layer) to produce a material consisting of layers of two or more different electrospun polymers. In certain embodiments, the electrospun material comprises nPET-PBT layered on nPU. In other embodiments, the electrospun material comprises nPU layered on nPET-PBT. Such a bilayer scaffold can be used to fabricate the cell chamber devices described herein.

[0086] One or more additional polymers can optionally be layered onto or between the aforementioned layers to form a multilayer material that can be used to fabricate some embodiments of the cell chamber devices described herein. In some embodiments, the multilayer scaffold can include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more layers of nanofibrous polymer, e.g., nanofibrous electrospun polymer. The layers can each include a different polymer, or multiple layers can include the same polymer.

[0087] The cell chamber can be fabricated from multi-layer (e.g., bi-layer) electrospun polymer material. The material can be measured and cut to any desired dimensions. Once prepared to the desired shape and size, the edges of the electrospun polymer material can be partially or completely sealed to form the chamber. The polymer material can be sealed at the edges of the chamber according to methods known in the art. In one embodiment, the edges of the chamber are sealed using ultrasonic welding. In some embodiments, at least one portion of the chamber is left unsealed, for example, to allow for subsequent cell loading. In some embodiments, the chamber is completely sealed but can later be opened, for example, by cutting or piercing with a needle, to create a cell loading port that can be subsequently sealed. In some embodiments, the scaffold material is folded before sealing the edges. In some embodiments, a cell loading port (e.g., a port with a luer lock mechanism that can be attached to a cell injection device) will be incorporated into the chamber membrane design to enable cell loading. After loading, the port is optionally removed and the chamber can be sealed by ultrasonic or heat welding, or by sealing the chamber opening with an adhesive.

[0088] In some cases, the nanofibrous polymer comprising the inner and / or outer layers of the scaffold of the cell chamber device herein can be further modified with one or more charged surface modifications. For example, in some embodiments, the nanofibrous polymer of the inner and / or outer layers of the scaffold is contacted with liquid ethylenediamine. Treatment with ethylenediamine, for example, when reacted with PET, results in a positively charged surface. In other embodiments, the nanofibrous polymer of the inner and / or outer layers of the scaffold is contacted with liquid sodium hydroxide. Treatment with PET, for example, when treated with sodium hydroxide, results in a negatively charged surface. Varying the surface charge of a material can be used to modulate the attraction or adhesion of cell types that have inherent charges on their cell surface proteins. Charged surface materials can also attract specific proteins, such as proteins in blood and the surrounding environment, which promote cell attachment. Ionic surfaces can also improve surface wettability and cell contact. Charged surface modifications may include cationic functional groups, anionic functional groups, or both cationic and anionic functional groups. For example, the charged surface modification can include carboxylic acid and / or amine functional groups. In some embodiments, the charged surface modification can be produced by treating the scaffold polymer with alkaline hydrolysis (e.g., by sodium hydroxide treatment). In some embodiments, the charged surface modification can be produced by treating the scaffold polymer with a diamine, such as ethylenediamine (EDA), 2-methylpentamethylenediamine, 1,2-diaminocyclohexane, or 1,6-hexanediamine. Examples of charged surface modifications and methods for producing charged surface modifications are further described, for example, in U.S. Patent Nos. 6,743,253 B2 and 7,037,527 B2, which are incorporated herein by reference in their entireties.

[0089] In some cases, the inner and / or outer scaffold layers of the cell chamber device further comprise a substance that reduces implant injury-related inflammation, such as a steroid (e.g., dexamethasone, triamcinilone) or an immunosuppressant (e.g., tacrolimus, sirolimus, everolimus, and / or paclitaxel). In some embodiments, the inner and / or outer scaffold layers of the cell chamber device comprise the cytokine CCL22, which can promote implant tolerance by recruiting Treg cells to the vicinity of the implant. The substance may be incorporated into the polymer during scaffold production or coated onto the scaffold after production. For example, the substance can be dissolved in the polymer solution before electrospinning. In such embodiments, the substance will be incorporated into the scaffold nanofibers during production. In other embodiments, the fully or partially formed scaffold can be immersed in a solution containing a substance that can be absorbed by the scaffold material. As described herein, introducing a net positive or net negative charge into the scaffold fibers can promote adhesion of substances loaded into the scaffold in this manner. Other substances, such as antibiotics, growth factors, etc., can be incorporated into the scaffold material in the manner described above.

[0090] The nanofibrous polymer scaffolds described herein can comprise pores resulting from the random and web-like arrangement of nanofibrous polymers. Additionally or alternatively, pores of any desired shape and diameter can be introduced into the scaffold material. In some embodiments, the pores are nanopores (e.g., having a size of 1 μm or less).

[0091] The pores can be sized to allow biomolecules secreted by cells in the chamber to pass out of the scaffold and into host tissue or blood, while preventing cells from passing into or out of the chamber. For example, the nanopores can be sized to allow the passage of biomolecules described herein, such as polypeptides, polysaccharides, and / or polynucleotides, as well as organic molecules, such as lipids (e.g., phospholipids, glycolipids, and sterols), vitamins, sugars (e.g., carbohydrates, disaccharides, oligosaccharides, polysaccharides), and / or amino acids. The pores can be sized to allow polypeptides (e.g., antibodies, hormones, neurotransmitters, etc.), peptides, oligopeptides, nucleotides, deoxyribonucleic acid (DNA), ribonucleic acid (RNA), or miRNA to diffuse across the scaffold. In certain embodiments, the pores are sized to allow antibodies, or antigen-binding portions thereof, to pass across the scaffold.

[0092] In some embodiments, the nanopore is sized to allow the passage of biomolecules having a molecular weight of about 250 kDa or less (e.g., about 225 kDa or less, about 220 kDa or less, about 200 kDa or less, about 175 kDa or less, about 150 kDa or less, about 125 kDa or less, about 100 kDa 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 nanopore is sized to allow the passage of biomolecules having a molecular weight of 10 kDa to 50 kDa, 50 kDa to 100 kDa, 100 kDa to 150 kDa, 130 kDa to 165 kDa, 150 kDa to 200 kDa, or 200 kDa to 250 kDa.

[0093] The nanopores may also be sized to prevent the passage of cells and / or prevent cells on one side of the scaffold from contacting cells on the other side of the scaffold. Thus, in some embodiments, the nanopores have a diameter of about 1 μm or less (e.g., about 0.9 μm or less, about 0.8 μm or less, about 0.7 μm or less, about 0.6 μm or less, about 0.5 μm or less, about 0.4 μm or less, about 0.3 μm or less, about 0.2 μm or less, about 0.1 μm or less, or about 0.05 μm or less). In certain embodiments, the nanopores have a diameter of 0.5 μm or less. In some embodiments, the nanopores have a diameter of 0.1 μm to 0.2 μm, 0.2 μm to 0.4 μm, 0.3 μm to 0.5 μm, 0.4 μm to 0.6 μm, 0.6 μm to 0.8 μm, or 0.8 μm to 1 μm. In some embodiments, the nanopore has a diameter of 0.1 μm to 3 μm. In other embodiments, the nanopore has a diameter of 0.2 μm to 2 μm. In some embodiments, the nanopore has a diameter of 1 nm to 10 μm (e.g., 1 nm to 10 nm, 5 nm to 25 nm, 10 nm to 50 nm, 25 nm to 75 nm, 50 nm to 100 nm, 75 nm to 125 nm, 100 nm to 200 nm, 150 nm to 250 nm, 200 nm to 300 nm, 250 nm to 500 nm, 300 nm to 400 nm, 350 nm to 450 nm, 400 nm to 500 nm, 450 nm to 550 nm, 500 nm to 600 nm). 00nm, 500nm to 1μm, 550nm to 650nm, 600nm to 700nm, 650nm to 750nm, 700nm to 800nm, 750nm to 850nm, 800nm ​​to 900nm, 900nm to 1μm, 1μm to 2μm, 1μm to 5μm, 1μm to 10μm, 2μm to 4μm, 2μm to 6μm, 2μm to 10μm, 4μm to 6μm, 4μm to 8μm, 4μm to 10μm, or 5μm to 10μm).

[0094] Antibodies generally have a length of approximately 10-15 nm. Many other peptide therapeutics are of comparable or smaller size. For example, Fab fragments have a length of approximately 9 nm. Therefore, pore diameters of approximately 10 nm, 15 nm, 20 nm, or greater are generally sufficient to allow peptides and antibodies to pass through the device, while diameters of approximately 10 μm or less (e.g., 9 μm or less, 8 μm or less, 7 μm or less, 6 μm or less, 5 μm or less, 4 μm or less, 3 μm or less, 2 μm or less, or 1 μm or less) are generally sufficient to retain encapsulated cells within the device. The exact size of the pores can be adjusted based on the size of the encapsulated cells and the size of the secreted biomolecules.

[0095] In some embodiments, the device may be a multi-scaffold device. A multi-scaffold device includes one or more nanofibrous polymer scaffolds in addition to the multi-layer scaffold described above. For example, the device may include (i) a first scaffold surrounding a cell chamber, the first scaffold including an outer layer and an inner layer, each of which includes a nanofibrous polymer; and (ii) a second scaffold positioned adjacent to the inner layer and in contact with the cell chamber. The second scaffold may optionally provide additional surface area for adhesion of cells within the cell chamber. In one embodiment, the second scaffold includes a nanofibrous polymer, e.g., an electrospun polymer. Any polymer described herein for incorporation into the first scaffold is similarly suitable for incorporation into the second scaffold. In an exemplary embodiment, the second scaffold may include nPET, nPBT, nPU, or a combination thereof, e.g., nPET-PBT. In an exemplary embodiment, the second scaffold includes nPBT. In some embodiments, the second scaffold comprises pores. Because the second scaffold is surrounded by the first scaffold, the pores can be of any size, for example, nanoporous or macroporous. In one embodiment, the second scaffold comprises pores of approximately the same size as the first scaffold. In another embodiment, the second scaffold comprises pores larger than those in the first scaffold. In another embodiment, the second scaffold comprises pores smaller than those in the first scaffold. An exemplary multi-scaffold device is shown in FIG. 1D.

[0096] In some embodiments, the outer (first) scaffold comprises pores of larger size than the pores of the inner (second) scaffold. For example, in some embodiments, the outer scaffold may comprise pores sized to allow capillary ingrowth for perfusion of the inner chamber bioreactor, while the inner scaffold comprises pores sized to allow the passage of biomolecules while blocking the passage of cells. In one embodiment, the outer scaffold contains pores of about 5-15 μm (e.g., 5-7 μm, 5-10 μm, 5-12 μm, 7-15 μm, 10-15 μm, etc.) that reflect the approximate diameter of a capillary, while the inner scaffold has a diameter of 1 μm or less (e.g., about 0.9 μm or less, about 0.8 μm or less, about 0.7 μm or less, about 0.6 μm or less, about 0.5 μm or less, about 0.4 μm or less, about 0.3 μm or less, about 0.2 μm or less, about 0.1 μm or less, or about 0.05 μm or less) to retain cells within the chamber.

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

[0098] In some embodiments, the membrane can comprise a nanofibrous polymer. For example, in some embodiments, the membrane can comprise nanofibrous polybutylene terephthalate. The nanofibrous membrane can contain pores resulting from the random web-like arrangement of the nanofibers, as described above with respect to the scaffold.

[0099] In other embodiments, the membrane is not nanofibrous. For example, the membrane can comprise a solid sheet interspersed with nanopores of a predetermined size. Non-nanofibrillar membranes can be composed of virtually any biocompatible polymer, including, but not limited to, any of the polymers described herein, such as PET, PBT, or PU. In some embodiments, non-nanofibrillar membranes can comprise one or more of the following polymers: mixed cellulose ester (MCE), cellulose acetate, coated cellulose acetate, hydrophilic PTFE, hydrophobic PTFE, nylon, or polycarbonate. In some embodiments, electrospun fibers can coat one or both sides of the membrane. For example, a solid (non-nanofibrillar) film containing pores of a defined size can be coated with electrospun fibers, such as nPET-PBT or nPU. In one embodiment, the scaffold comprises a solid (non-nanofibrillar) membrane interspersed with nanopores, coated with electrospun nPET-PBT on one side and electrospun nPU on the other side. The nanopore-studded solid membrane can comprise a non-nanofibrillar polymer sheet (e.g., comprised of PET, PBT, PU, ​​or other suitable polymers, or a combination thereof). In an exemplary embodiment, the solid membrane comprises PET. In another exemplary embodiment, the solid membrane comprises PU. Incorporating a non-nanofibrillar membrane into the scaffold can act as a barrier to block the passage of cells that would otherwise be forced through the nanofibrous material. In such an embodiment, the membrane can comprise pores sized to block the passage of cells. Scaffolds containing a solid (non-nanofibrillar) membrane as their outer surface can cause inflammation in host subjects at the implantation site. Coating a solid (non-nanofibrillar) membrane with electrospun fibers can significantly reduce the inflammatory response to the scaffold compared to a scaffold comprised of a solid membrane without a coating of electrospun fibers.

[0100] The membrane nanopore can be sized to allow the passage of biomolecules, e.g., proteins, across the membrane while blocking the passage of cells. In one embodiment, the membrane pore is sized to allow the passage of biomolecules having a molecular weight of about 250 kDa or less (e.g., about 225 kDa or less, about 220 kDa or less, about 200 kDa or less, about 175 kDa or less, about 150 kDa or less, about 125 kDa or less, about 100 kDa 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 pore is sized to allow the passage of biomolecules having a molecular weight of 10 kDa to 50 kDa, 50 kDa to 100 kDa, 100 kDa to 150 kDa, 150 kDa to 200 kDa, or 200 kDa to 250 kDa. The membrane pores may also be sized to prevent the passage of cells and / or prevent cells on one side of the scaffold from contacting cells on the other side of the scaffold. Thus, in some embodiments, the membrane comprises nanopores having a diameter of about 1 μm or less (e.g., about 0.9 μm or less, about 0.8 μm or less, about 0.7 μm or less, about 0.6 μm or less, about 0.5 μm or less, about 0.4 μm or less, about 0.3 μm or less, about 0.2 μm or less, about 0.1 μm or less, or about 0.05 μm or less). In certain embodiments, the membrane nanopores have a diameter of 0.5 μm or less. In some embodiments, the membrane nanopores have a diameter of 0.1 μm to 0.4 μm, 0.4 μm to 0.6 μm, 0.6 μm to 0.8 μm, or 0.8 μm to 1 μm. In some embodiments, the membrane nanopores have a diameter of 0.1 to 3 μm. In other embodiments, the membrane nanopores have a diameter of 0.2 to 2 μm.In some embodiments, the membrane nanopores are between 1 nm and 10 μm (e.g., 1 nm to 10 nm, 5 nm to 25 nm, 10 nm to 50 nm, 25 nm to 75 nm, 50 nm to 100 nm, 75 nm to 125 nm, 100 nm to 200 nm, 150 nm to 250 nm, 200 nm to 300 nm, 250 nm to 500 nm, 300 nm to 400 nm, 350 nm to 450 nm, 400 nm to 500 nm, 450 nm to 550 nm, 500 nm to The membrane pores have a diameter of 600 nm, 500 nm to 1 μm, 550 nm to 650 nm, 600 nm to 700 nm, 650 nm to 750 nm, 700 nm to 800 nm, 750 nm to 850 nm, 800 nm to 900 nm, 900 nm to 1 μm, 1 μm to 2 μm, 1 μm to 5 μm, 1 μm to 10 μm, 2 μm to 4 μm, 2 μm to 6 μm, 2 μm to 10 μm, 4 μm to 6 μm, 4 μm to 8 μm, 4 μm to 10 μm, or 5 μm to 10 μm. In some embodiments, the membrane pores are approximately the same size as the pores in the outer and / or inner layers of the nanofibrous polymer scaffold. In other 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 larger than the pores in the outer and / or inner layers of the nanofibrous polymer scaffold.

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

[0102] Like the inner or outer layer of the scaffold, the nanoporous film or membrane may comprise membrane nanopores sized to allow the passage of biomolecules secreted by cells within the cell across the scaffold into the host tissue or blood, and / or to allow the passage of biomolecules (e.g., nutrients) from the host into the chamber to supply cells seeded on the device. For example, the membrane nanopores may be sized to allow the passage of any of the biomolecules described herein, e.g., polypeptides, polysaccharides, and polynucleotides, as well as organic molecules such 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 acid (DNA), or ribonucleic acid (RNA). In certain embodiments, the membrane nanopores are sized to allow the passage of proteins, e.g., antibodies, or antigen-binding portions thereof.

[0103] In some embodiments, the membrane nanopore is sized to allow the passage of biomolecules having a molecular weight of about 250 kDa or less (e.g., about 225 kDa or less, about 220 kDa or less, about 200 kDa or less, about 175 kDa or less, about 150 kDa or less, about 125 kDa or less, about 100 kDa 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 nanopore is sized to allow the passage of biomolecules having a molecular weight of 10 kDa to 50 kDa, 50 kDa to 100 kDa, 100 kDa to 150 kDa, 130 to 165 kDa, 150 kDa to 200 kDa, or 200 kDa to 250 kDa.

[0104] In an exemplary embodiment, the cell chamber device includes a multi-layer scaffold surrounding the cell chamber, the multi-layer scaffold including an outer layer including nanofibrous polyethylene terephthalate and polybutylene terephthalate and an inner layer including nanofibrous polyurethane, the outer layer and the inner layer having a thickness of 1 nm to 10 μm (e.g., 1 nm to 10 nm, 5 nm to 25 nm, 10 nm to 50 nm, 25 nm to 75 nm, 50 nm to 100 nm, 75 nm to 125 nm, 100 nm to 200 nm, 150 nm to 250 nm, 200 nm to 300 nm, 250 nm to 500 nm, 300 nm to 400 nm, Nanopores with diameters of 00nm, 350nm to 450nm, 400nm to 500nm, 450nm to 550nm, 500nm to 600nm, 500nm to 1µm, 550nm to 650nm, 600nm to 700nm, 650nm to 750nm, 700nm to 800nm, 750nm to 850nm, 800nm ​​to 900nm, 900nm to 1µm, 1µm to 2µm, 1µm to 5µm, 1µm to 10µm, 2µm to 4µm, 2µm to 6µm, 2µm to 10µm, 4µm to 6µm, 4µm to 8µm, 4µm to 10µm, or 5µm to 10µm.In another exemplary embodiment, the cell chamber device includes a three-layer scaffold surrounding the cell chamber, the three-layer scaffold including an outer layer including nanofibrous polyethylene terephthalate and polybutylene terephthalate, an inner layer including nanofibrous polyurethane, and a membrane positioned between the inner and outer layers, the inner layer including nanofibrous polybutylene terephthalate, wherein the outer layer, the inner layer, and the membrane are sized to a thickness of 1 nm to 10 μm (e.g., 1 nm to 10 nm, 5 nm to 25 nm, 10 nm to 50 nm, 25 nm to 75 nm, 50 nm to 100 nm, 75 nm to 125 nm, 100 nm to 200 nm, 150 nm to 250 nm, 200 nm to 300 nm). Nanopores with diameters of 0 nm, 250 nm to 500 nm, 300 nm to 400 nm, 350 nm to 450 nm, 400 nm to 500 nm, 450 nm to 550 nm, 500 nm to 600 nm, 500 nm to 1 μm, 550 nm to 650 nm, 600 nm to 700 nm, 650 nm to 750 nm, 700 nm to 800 nm, 750 nm to 850 nm, 800 nm to 900 nm, 900 nm to 1 μm, 1 μm to 2 μm, 1 μm to 5 μm, 1 μm to 10 μm, 2 μm to 4 μm, 2 μm to 6 μm, 2 μm to 10 μm, 4 μm to 6 μm, 4 μm to 8 μm, 4 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 of any size or shape suitable for administration to a host subject (e.g., administration via surgery such as minimally invasive surgery, e.g., laparoscopic or endoscopic surgery). The size and shape can be modified as needed to accommodate the desired number of cells that can be loaded into the device, to adapt the device to the implantation site, and / or to regulate the dosage of therapeutic biomolecules secreted by cells within the device to the recipient subject (e.g., as measured by mg of biomolecule secreted from the device per day).

[0107] In some embodiments, the cell chamber contains up to about 1 x 10 11 cells (e.g., up to approximately 1 x 1011 cells, up to approximately 1 x 10 10 cells, up to approximately 1 x 10 9 cells, up to approximately 1 x 10 8 cells, up to approximately 1 x 10 7 cells, up to approximately 1 x 10 6 cells, up to approximately 1 x 10 5 cells, up to approximately 1 x 10 4 cells, or up to approximately 1 x 10 3 In some embodiments, the cell chamber is sized to accommodate up to 1 x 10 cells. 7 In some embodiments, the cell chamber is sized to accommodate up to 1 x 10 cells. 11 The chamber is sized to accommodate 1 x 10 cells. The number of cells to be loaded into the device can be easily determined by one skilled in the art based on the desired dose of biomolecule to be delivered and the amount of biomolecule produced by the cells. For example, if a 10 μg dose per day is required and the cells produce 1 pg / cell / day, the chamber should accommodate 1 x 10 cells. 7 The cell can be designed to accommodate a number of cells.

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

[0109] In certain embodiments, the cell chamber device is rectangular. For example, in some embodiments, the rectangular cell chamber device can have a length of about 15 cm or less, about 12 cm or less, about 10 cm or less, about 8 cm or less, about 6 cm or less, about 4 cm or less, about 2 cm or less, about 1 cm or less, about 0.1 cm (100 mm) or less, or about 0.01 cm (10 mm) or less. In some embodiments, the rectangular cell chamber device has a width of about 15 cm or less, about 12 cm or less, about 10 cm or less, about 8 cm or less, about 6 cm or less, about 4 cm or less, about 2 cm or less, about 1 cm or less, about 0.1 cm (100 mm) or less, or about 0.01 cm (10 mm) or less. In some embodiments, the cell chamber device has a width of about 5-10 cm and a length of about 10-15 cm. In some embodiments, the cell chamber device has a width of about 3-5 cm and a length of about 5-10 cm. In some embodiments, the cell chamber device has a width of about 1-3 cm and a length of about 3-5 cm. In some embodiments, the cell chamber device has a width of about 0.01-1 cm and a length of about 1-3 cm. In some embodiments, the cell chamber device has a first dimension of about 0.01 cm, about 0.1 cm, about 1 cm, about 2 cm, about 3 cm, about 4 cm, about 5 cm, about 6 cm, about 7 cm, about 8 cm, about 9 cm, about 10 cm, about 11 cm, about 12 cm, about 13 cm, about 14 cm, or about 15 cm, and a second dimension of about 0.01 cm, about 0.1 cm, about 1 cm, about 2 cm, about 3 cm, about 4 cm, about 5 cm, about 6 cm, about 7 cm, about 8 cm, about 9 cm, about 10 cm, about 11 cm, about 12 cm, about 13 cm, about 14 cm, or about 15 cm. In certain embodiments, the cell chamber device has a width of about 8 cm and a length of about 10 cm. In other embodiments, the cell chamber device has a width of about 5 cm and a length of about 9 cm. In other embodiments, the cell chamber device has a width of about 3 cm and a length of about 5 cm. In other embodiments, the cell chamber device has a width of about 1 cm and a length of about 3 cm.

[0110] In some embodiments, the device is about 250 cm 2 For example, about 225 cm 2 Below, approximately 220cm2 Below, approximately 200cm 2 Below, approximately 175cm 2 Below, approximately 150cm 2 Below, approximately 125cm 2 Below, approximately 100cm 2 Below, approximately 75cm 2 Below, approximately 50cm 2 Below, approximately 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, approximately 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 comprises a total thickness that is no greater than 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% of the next smallest dimension of the device, e.g., width. In some embodiments, the device comprises a total thickness of about 1 mm or less, 0.9 mm or less, 0.8 mm or less, 0.7 mm or less, 0.6 mm or less, 0.5 mm or less, 0.4 mm or less, 0.3 mm or less, 0.2 mm or less, or 0.1 mm or less. In some embodiments, the device comprises a total thickness of 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 μm or less, about 50 μm or less, about 25 μm or less, or about 10 μm or less). In some embodiments, the device has a diameter of 10 μm to 50 μm, 10 μm to 100 μm, 50 μm to 100 μm, 50 μm to 200 μm, 100 μm to 150 μm, 100 μm to 250 μm, 100 μm to 500 μm, 150 μm to 200 μm, 200 μm to 250 μm, 250 μm to 300 μm, 250 μm to 500 μm, 300 μm to 350 μm, 350 μm to 400 μm, 40 In certain embodiments, the device comprises a total thickness of 0 μm to 450 μm, 450 μm to 500 μm, 500 μm to 550 μm, 500 μm to 1 mm, 550 μm to 600 μm, 600 μm to 650 μm, 650 μm to 700 μm, 700 μm to 750 μm, 750 μm to 800 μm, 750 μm to 1 mm, 800 μm to 850 μm, 850 μm to 900 μm, 900 μm to 950 μm, or 950 μm to 1 mm. In certain embodiments, the device comprises a total thickness of 150 μm or less.

[0113] The cell chamber devices disclosed herein may optionally include a loading port to facilitate cell loading into the cell chamber. Such a loading port can traverse the layers of the multilayer scaffold. The loading port can include an external opening facing the outside of the cell chamber device (e.g., for inserting a cell loading or injection instrument) and an internal opening facing the inside of the cell chamber (e.g., for depositing cells into the cell chamber via a cell loading or injection instrument). The loading port can be sealed to prevent the loaded cells in the cell chamber from leaking out of the cell chamber. After cell loading, the opening in the chamber can be sealed by various methods, such as ultrasonic welding. Other examples of sealing processes known in the art include heat staking (e.g., cold staking or heat staking), swaging, spin welding, hot plate welding, vibration welding, or laser welding.

[0114] III.Cells The devices disclosed herein can accommodate cells in a cell chamber surrounded by a multilayer scaffold. Thus, in some embodiments, the device can optionally contain cells. Cells can be loaded into the cell chamber through a cell loading port or through an opening in the bilayer scaffold, which can then be sealed, for example, using the methods described herein. The cell chamber is contacted by the inner layer of the bilayer scaffold so that the cells loaded into the cell chamber can optionally adhere to the inner layer of the scaffold.

[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 robust under harsh conditions, (2) the cells can be genetically engineered to produce (e.g., secrete) a desired therapeutic biomolecule, (3) the cells can have a relatively long lifespan or shelf life (e.g., greater than one month), (4) if the subject is human, the cells can be of human origin to increase compatibility between the encapsulated cells and the host, (5) the cells can exhibit high viability in the device to ensure long-term delivery (e.g., greater than 80% viability in vivo within the device for a period of greater than one month), (6) the encapsulated cells can deliver effective amounts of a useful biological product, (7) the cells can have a low level of host immune response to ensure longevity of the implant, and / or (8) the cells can be non-tumorous to provide additional safety in the event of device leakage.

[0116] In some embodiments, mammalian cells known in the art for the production of mammalian proteins may be suitable for use in the cell chamber device. Chinese hamster ovary (CHO) cells, as well as cell lines derived from various other mammalian sources, such as mouse myeloma (NS0), baby hamster kidney (BHK), human embryonic kidney (HEK-293), and human retinal cells, have been approved by regulatory agencies for the production of biopharmaceuticals, including therapeutic antibodies. Examples of mammalian host cells include CHO, BHK, HEK293, COS, PC12, HiB5, RN33b, C2C12, HepG2, and ARPE-19 cells.

[0117] In certain embodiments, the cells are human cells, including recombinant cells of human origin. In one embodiment, the cells comprise human retinal pigment epithelial (RPE) cells or cells derived therefrom. In an exemplary embodiment, the cells comprise ARPE-19 cells. The ARPE-19 cell line (see, e.g., Dunn et al., 62 Exp. Eye Res. 155-69 (1996); Dunn et al., 39 Invest. Ophthalmol. Vis. Sci. 2744-9 (1998); Finnemann et al., 94 Proc. 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); U.S. Patent No. 6,361,771) possesses many characteristics of a platform cell line for use with the cell chamber devices described herein. The ARPE-19 cell line is available from the American Type Culture Collection (ATCC No. CRL-2302). ARPE-19 cells are normal retinal pigment epithelial (RPE) cells and express the retinal pigment epithelial cell-specific markers CRALBP and RPE-65. ARPE-19 cells form stable monolayers that exhibit morphological and functional polarity. The ARPE-19 cell line is viable under harsh conditions, such as during implantation into a host subject, can be genetically engineered to secrete biomolecules of therapeutic interest, has a relatively long lifespan, and is human-derived. Additionally, encapsulated ARPE-19 cells have good in vivo survival in devices, elicit a non-significant immune response in human hosts, and are not tumorigenic. In another embodiment, the cells are human hepatocytes. In another embodiment, the cells are human pancreatic islet cells.

[0118] The cells can be transformed or non-transformed. Furthermore, the cells can be grown in suspension or in an adherent state. In some embodiments, the cells are contact-inhibited cells, e.g., immortalized contact-inhibited human cells (e.g., the hTERT immortalized cell line (Evercyte, Vienna, AT). Cells are known in the art for recombinant protein production. For example, mammalian cell lines that can be used in the devices described herein include, in some embodiments, SV40-transformed monkey kidney CVI line (COS-7, ATCC™ CRL1651), baby hamster kidney cells (BHK, ATCC™ CCL10), mouse Sertoli cells (TM4, Mather, Biol. Reprod., 23:243 (1980)), monkey kidney cells (CVI-76, ATCC™ CCL70), African green monkey kidney cells (VERO-76, ATCC™ CRL-1587), dog kidney cells (MDCK, ATCC™ CCL34), buffalo rat liver cells (BRL3A, ATCC.RTM.CRL1442), mouse mammary carcinoma cells (MMT060562, ATCC V CCL51), rat hepatocytes (HTC, MI.54, Baumann et al., J. Cell Biol., 85:1 (1980)), 3T3 cells, 293T cells (Pear, W.S., 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., Annals NY Acad. Sci., 383:44 (1982)), and hybridoma cell lines.In some embodiments, the cell line is a human cell line such as the human embryonic kidney line 293S (Graham et al., J. Gen. Virol., 36:59 (1977)), human cervical carcinoma cells (HELA, ATCC™ CCL2), human lung cells (W138, ATCC™ CCL75), human liver cells (Hep G2.HB8065), an hTERT-immortalized cell line (Evercyte, Vienna, AT), or human retinal cells. In some embodiments, the cell line is a carcinogenic cell line that has been modified or treated to make it safe for use in the cell chamber devices provided herein. In some embodiments, the cell chamber device can contain stem cells, e.g., human stem cells such as induced pluripotent stem cells (iPS), 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 device of the present disclosure may be genetically modified cells, e.g., recombinant cells, engineered to produce, e.g., secrete, a biomolecule of interest. Biomolecules that may be secreted by the cells of the device include, but are not limited to, polypeptides, polysaccharides, and polynucleotides, as well as organic molecules such as lipids (e.g., phospholipids, glycolipids, and sterols), chemical messengers (e.g., neurotransmitters and hormones such as insulin), vitamins, sugars (e.g., carbohydrates, disaccharides, oligosaccharides, and polysaccharides), amino acids, peptides, oligopeptides, polypeptides, proteins, nucleotides, deoxyribonucleic acid (DNA), or ribonucleic acid (RNA). Other secreted biomolecules may include those assembled, packaged, and secreted as exosomes, lipid polymers, or viral particles. In some embodiments, the cells produce therapeutic biomolecules as described herein.

[0120] In some embodiments, the cells in the device can be engineered to secrete one or more proteins or peptides, e.g., one or more recombinant proteins or peptides, e.g., one or more therapeutic proteins or peptides. For example, the cells in the device can secrete one or more therapeutic proteins, such as an antibody or antigen-binding fragment thereof, a growth factor, a hormone (e.g., insulin), a cytokine, a clotting factor (e.g., Factor VIII or Factor IX, or variants thereof, e.g., Recombinate, Kogenate, Refacto, Advate, Alprolix, BeneFIX, Rixubis, Ixinity, Idelvion, etc.), or a combination thereof. The protein can, in some cases, be a recombinant protein or peptide.

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

[0122] In some embodiments, the cells within the device are capable of secreting one or more growth factors, including, but not limited to, fibroblast growth factor (FGF), epidermal growth factor (EGF), platelet-derived growth factor (PDGF), insulin-like growth factor (IGF), transforming growth factor (TGF), vascular endothelial growth factor (VEGF), liver growth factor (LGF), bone morphogenetic protein (BMP), colony-stimulating factor (CSF), hepatocyte growth factor (HGF), or nerve growth factor (NGF), or a combination thereof.

[0123] In some embodiments, the cells within the device secrete one or more cytokines, including, but not limited to, bone morphogenetic protein (BMP), erythropoietin (EPO), granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon alpha, interferon beta, interleukin 2 (IL-2), interleukin 11 (IL-11), or interferon gamma, or a combination thereof.

[0124] In some embodiments, the cells within the device secrete one or more hormones, including, but not limited to, insulin, estrogen, progestogen, thyroxine (as levothyroxine), or steroids, or combinations thereof.

[0125] In some embodiments, the cells in the device secrete one or more enzymes. For example, the cells in the device can secrete an enzyme that provides enzyme replacement therapy (ERT) to a subject. In some embodiments, the cells can produce an enzyme that complements lysosomal storage disease deficiencies, including, but not limited to, hexosaminidase A, alpha-galactosidase A, glucocerebrosidase, arylsulfatase A, galactocerebrosidase, and sphingomyelinase. In some embodiments, the cells can produce an enzyme that complements Hunter syndrome, also known as mucopolysaccharidosis type II (MPSII), including, but not limited to, idursulfase. In some embodiments, the cells can produce an enzyme that complements metachromatic leukodystrophy (MLD), including, but not limited to, arylsulfatase A. In some embodiments, the cells can produce an enzyme that complements metachromatic leukodystrophy (MLD), including, but not limited to, laronidase. It can produce enzymes that complement mucopolysaccharidosis type I (MPSI).

[0126] In some embodiments, cells within the device are capable of secreting one or more antibodies, or antigen-binding portions thereof. The antibodies, or antigen-binding portions thereof, described herein may be in the form of full-length antibodies, bispecific antibodies, dual variable domain antibodies, multi- or single-chain antibodies, and / or antigen-binding fragments that specifically bind to extracellular molecules, including, but not limited to, Fab, Fab', (Fab')2, Fv, scFv (single-chain Fv), surrobodies (including surrogate light chain constructs), single-domain antibodies, camelized antibodies, etc. They may also be of or derived from any isotype, including, for example, IgA (e.g., IgA1 or IgA2), IgD, IgE, IgG (e.g., IgG1, IgG2, IgG3, or IgG4), or IgM. In some embodiments, the cells secrete antibody fragments selected from the group consisting of Fab, F(ab')2, scFv, tandem scFv, diabodies, minibodies, and single-domain antibodies. In some embodiments, the cells secrete full-length antibodies.

[0127] In some embodiments, the cells secrete chimeric antibodies, or antigen-binding portions thereof. The term "chimeric antibody" is intended to refer to antibodies in which the variable region sequences are derived from one species and the constant region sequences are derived from another species, e.g., antibodies in which the variable region sequences are derived from a murine antibody and the constant region sequences are derived from a human antibody.

[0128] In some embodiments, the cells secrete the humanized antibody, or an antigen-binding portion thereof. "Humanized" forms of non-human (e.g., rodent) antibodies are chimeric antibodies that contain minimal sequence derived from the non-human antibody. For the most part, humanized antibodies are human immunoglobulins (recipient antibodies) in which residues from a hypervariable region of a human "recipient" antibody are replaced by residues from a hypervariable region of a "donor" antibody from a non-human species, such as mouse, rat, rabbit, or non-human primate, having the desired specificity, affinity, and capacity. In some cases, framework region (FR) residues of the human antibody may be replaced by corresponding non-human residues. Furthermore, in some cases, humanized antibodies can comprise residues that are not found in either the recipient or donor antibody. These modifications can further improve antibody performance. In some embodiments, humanized antibodies comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable CDR loops correspond to the hypervariable loops of a non-human antibody and all or substantially all of the FRs are those of a human antibody sequence. The humanized antibody optionally also will comprise at least a portion of an antibody constant region (Fc), typically that of a human antibody. For further details, see, e.g., Jones et al., Nature 321:522-525 (1986), Riechmann et al., Nature 332:323-329 (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 "human antibody," as used herein, refers to antibodies having variable regions in which both the framework and CDR regions are derived from human germline immunoglobulin sequences. Furthermore, if the antibody contains a constant region, the constant region also is derived from human germline immunoglobulin sequences. The human antibodies of the invention may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). However, the term "human antibody," as used herein, is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences.

[0130] In some embodiments, the cells secrete monoclonal antibodies. As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical in sequence and specificity, but excludes variant antibodies that contain, for example, naturally occurring mutations or that may arise during the production of a monoclonal antibody preparation; such variants are generally present in minor amounts. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on an antigen. Thus, the modifier "monoclonal" indicates the character of the antibody as being obtained from a population of substantially homogeneous antibodies and is not to be construed as requiring isolation of the antibody by any particular method. For example, monoclonal antibodies to be used in accordance with the present invention may be derived using a variety of techniques, including, but not limited to, hybridoma methods, recombinant DNA methods, phage display methods, and methods utilizing transgenic animals containing all or portions of the human immunoglobulin loci; such methods and other exemplary methods for making monoclonal antibodies are known in the art and are described herein.

[0131] If the cells in the cell chamber secrete an antibody, or antigen-binding portion thereof, the antibody, or antigen-binding portion thereof, can specifically bind to any antigen of interest. In some embodiments, the antibody, or antigen-binding portion thereof, specifically binds to (i) a tumor-associated antigen, (ii) a cell surface receptor, (iii) CD proteins and their ligands, e.g., CD3, CD4, CD8, CD19, CD20, CD22, CD25, CD32, CD33, CD34, CD40, CD44, CD47, CD54, CD59, CD70, CD74, CD79a (CD79a), and CD79P (CD79b), (iv) a member of the ErbB receptor family, e.g., EGF receptor, HER2, HER3, or HER4 receptor. receptors, (v) cell adhesion molecules, such as LFA-1, Mac1, p150,95, VLA-4, ICAM-1, VCAM, and αv / β3 integrin containing either the alpha or beta subunits (e.g., anti-CD11a, anti-CD18, or anti-CD11b antibodies), or (vi) growth factors, such as PDGF, FGF, VEGF, IgE, blood group antigens, flk2 / flt3 receptor, obesity (OB) receptor, mpl receptor, CTLA4, protein C, BR3, c-met, tissue factor, and the like. In other exemplary embodiments, the antibody, or antigen-binding portion thereof, is selected from the group consisting of 4-1BB, 5T4, ACVR2B, ADAM-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, CD4, 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, CD166, CD184, CD200, CD205, CD221, CD248, CD276,CD278, CD279, CD319, CD352, CDH-6, CEA, CEA-CAM4, CEA-CAM5, CEA-Cide, CEA-related antigen, CEACAM1, CEACAM6, CFC1B, claudin 18 isoform 2, CLDN6, CLDN18.2, CSF1R, CTLA4, CXCR4, dendritic cell-associated lectin 2, DLK1, DLL3, DLL4, DR5, EFNA4, EGFR, EGFR extracellular domain III, EGFRviii, endoglin, endothelin receptor ETB, ENPP3, EpCAM, EphA, EPHA 3, ephrin receptor A3, ephrin A4, episialin, ER-alpha 36, ​​ERBB1, ERBB2, ERBB3, FCGRT, FGFR, FGFR2, fibronectin extra domain-B, FLT3, folate receptor, folate receptor 1, folate receptor alpha, FOLHI, Frizzled receptor, FXYD5, ganglioside GD3, GCC, GCGR, GCL, GD2 ganglioside, GD3 ganglioside, globo H, glypican 3, GMCSF receptor α chain, GPC2, GPNMB, granulocyte antigen, GUCY2C, H-ferritin, hepatitis B Inflammatory surface antigen, HER1, HER2, HER2 / neu, HER3, HGFR, HLA-DR, human scatter factor receptor kinase, ICAM-1, ICOS, IgE receptor, IGF1R, IL6R, IL31RA, IL3 receptor, IL-4Rα, IL-6R, IL-12 / 23, IL-17 receptor, integrin α4, integrin α4β7, integrin α5β1, integrin αIIbβ3, integrin αvβA, integrin β7, interferon receptor, interferon α / β receptor, ITGA2, 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, MT1-MMP, MTX3, MTX5, MUC1, mucin 16, mucosal addressin cell adhesion molecule (MAdCAM), myelin-associated glycoprotein, NCA-90, nectin-4, NGNA ganglioside, NKA, NKG2A, Notch3, Notch1, Notch receptor, NRP1, OFP, NaPi2b, OX-40, P-cadherin,P. aeruginosa antigen, PCDC1, PD-1, PD-L1, PDCD1, PDGF-Rα, PDGFRA, sodium phosphate cotransporter, phosphatidylserine, platelet-derived growth factor receptor beta, PRLR, PSMA, PTK7, RGMA, RHD, rhesus factor, rhesus factor, ROR1, Ror2, RSVFR, SAIL, SDC1, selectin P, SLAMF7, SLC34A2, SLC44A4, SLeA, SLITRK5, SLITRK6, Antibodies capable of specifically binding to antibodies include, but are not limited to, soluble IL-6, SOST, SSTR2, STEAP1, STn, T cell receptor, TACSTD2, 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. Other examples of antigens that can be targeted by antibodies or antigen-binding fragments thereof include cell surface receptors such as those described in Chen and Flies, Nature Reviews Immunology, 13.4 (2013):227, which is incorporated herein by reference.

[0132] In some embodiments, antibodies secreted by cells in the device include 3F8 (binds to GD2 ganglioside), abciximab (ReoPro; binds to CD41), abituzumab (binds to CD51), alemtuzumab (Lemtrada, Campath; binds to CD52), abrilumab (binds to integrin α4β7), adalimumab (Humira; binds to TNF-α), adecatumumab (binds to EpCAM), alacizumab pegol (binds to VEGFR2), acridumab (binds to EpCAM), acrid ... Lemtuzumab (Lemtrada, Campath; binds to CD52), altumomab pentetate (Hybri-ceaker; binds to CEA), amatuximab (binds to mesothelin), anatumomab mafenatox (binds to TAG-72), anetumab ravtansine (binds to MSLN), anifrolumab (binds to interferon α / β receptor), apolizumab (binds to HLA-DR), apolizumab ixadotin (binds to FGFR2), arcitumomab (binds to CEA) ), acelizumab (binding to L-selectin or CD62L), atezolizumab (Tecentriq; binding to PD-L1), atrolimumab (binding to rhesus factor), avelumab (Bavencio; binding to PD-L1), abicixizumab (DLL4; binding to VEGFA), azintuxizumab vedotin (binding to CD319), basiliximab (Simulect; binding to CD25), bavituximab (binding to phosphatidylserine), BCD-100 ( PD-1), vectamumomab (LymphoScan; binds to CD22), belantamab mafodotin (binds to BCMA), belimumab (Benlysta; binds to BAFF), bemarituzumab (binds to FGFR2), benralizumab (Fasenra; binds to CD125), velsanlimab (binds to ICAM-1), besilesomab (Scintimun; binds to CEA-related antigen), bimagrumab (binds to ACVR2B), bivatuzumab mertansine (CD44 v6), bleselumab (binds to CD40), blinatumomab (Blincyto; binds to CD19), brosozumab (binds to SOST), brentuximab vedotin (Adcentris; binds to CD30), brontuzumab (binds to Notch1), brodalumab (Siliq;IL-17 receptor), cabilalizumab (CSF1R), camidanlumab tesirin (CD25), camrelizumab (PD-1), carotuximab (endoglin), catumaxomab (Removab; EpCAM / CD3), cantuzumab ravtansine (MUC1), caplacizumab (Cablivi; VWF), cedelizumab (CD4), semipilimab (Libtayo; PCDC1), and ceto Relimab (binds to PD-1), certolizumab (binds to TNF-α), sergituzumab amnalikine (binds to CEA), cetuximab (Erbitux; binds to EGFR), sibisatamab (binds to CEACAM5), cirumutuzumab (binds to ROR1), cixutumumab (IGF-1 receptor; binds to CD221), clenoliximab (binds to CD4), clivatuzumab tetraxetan (hPAM4-Cide; binds to MUC1), codrituzumab (glypican 3), coltuximab ravtansine (binds to CD19), conatumumab (binds to TRAIL-R2), crizanlizumab (binds to selectin P), clotidumab (binds to GCGR), dacetuzumab (binds to CD40), daclizumab (Zenapax; binds to CD25), dalotuzumab (IGF-1 receptor; binds to CD221), dapirolizumab pegol (binds to CD154; binds to CD40L), daratumumab (Darzalex; binds to CD38), dem Tucizumab (binding to DLL4), denintuzumab mafodotin (binding to CD19), depatuxizumab mafodotin (binding to EGFR), drozizumab (binding to DR5), DS-8201 (binding to HER2), deligotuzumab (ERBB3; binding to HER3), dinutuximab (Unituxin; binding to GD2 ganglioside), dupilumab (binding to IL-4Rα), durvalumab (Imfinzi; binding to PD-L1), duvoltuximab (CD19;CD3E), ecloneximab (binding to GD3 ganglioside), edrecolomab (binding to EpCAM), elezanumab (binding to RGMA), elgemuzumab (binding to ERBB3, HER3), elotuzumab (binding to SLAMF7), emactuzumab (binding to CSF1R), enapotamab vedotin (binding to AXL), enavatuzumab (binding to TWEAK receptor), enri Enlimonomab pegol (ICAM-1; binds to CD54), enoblitzumab (binds to CD276), enoticumab (binds to DLL4), epratuzumab (binds to CD22), erlizumab (ITGB2; binds to CD18), ertumakizumab (Rexomun; binds to HER2 / neu; binds to CD3), etaracizumab (Abergin; integrin α; vβ3), etigilimab (binding to TIGIT), etrolizumab (binding to integrin β7), exbivirumab (binding to hepatitis B surface antigen), fanolesomab (binding to NeutroSpec; binding to CD15), faralimomab (binding to interferon receptor), farletuzumab (binding to folate receptor 1), FBTA05 (Lymphomun; binding to CD20), fugatipotuzumab (binding to MUC1), fivatuzumab (binding to ephrin receptor A3) ), figitumumab (IGF-1 receptor; binds to CD221), flotetuzumab (IL3 receptor; binds to CD3 epsilon), futuximab (binds to EGFR), galiximab (binds to CD80), gancotamab (binds to HER2 / neu), ganitumab (IGF-1 receptor; binds to CD221), gavilimomab (CD147; binds to basigin), gemtuzumab ozogamicin (Mylotarg; binds to CD33), glenbatumumab vedotin (binds to GPNMB), golimumab ( Simponi (binds to TNF-α), gomiliximab (CD23 (binds to IgE receptor)), ianalumab (binds to BAFF-R), ibalizumab (Trogarzo (binds to CD4)), IBI308 (binds to PD-1), ibritumomab tiuxetan (binds to CD20), icrucumab (binds to VEGFR-1), ifavotuzumab (binds to EPHA3), igovomab (Indimacis-125 (binds to CA-125)), IMAB362 (binds to CLDN18.2), imaprelimab (binds to MCAM), Inclacumab (binds to selectin P), indatuximab ravtansine (binds to SDC1), iradatuzumab vedotin (binds to CD97B), imgatuzumab (binds to EGFR), indusatumab vedotin (binds to GUCY2C), inebilizumab (binds to CD19), infliximab (Remicade; binds to TNF-α), intetumumab (binds to CD51), inolimomab (binds to CD25), inotuzumab ozogamicin (Besponsa; binds to CD22), ipilimumab (Yervoy;binds to CD152), Iomab-B (binds to CD45), Iratumumab (binds to CD30), Isatuximab (binds to CD38), Iscalimab (binds to CD40), Istiratumab (IGF1R; binds to CD221), Itolizumab (Alzumab; binds to CD6), Keliximab (binds to CD4), Laprituximab Emtansine (binds to EGFR), Labetuzumab (CEA-Cide; binds to CEA), Rifastuzumab Vedotin (binds to sodium phosphate cotransporter), Remaresomab (NCA-90; binds to granulocyte antigen), Lembervimab (binding to hepatitis B surface antigen), leronlimab (binding to CCR5), lexatumumab (binding to TRAIL-R2), ribivirumab (binding to hepatitis B surface antigen), loncusximab tesirin (binding to CD19), rosatuxizumab vedotin (binding to EGFR; ERBB1; HER1), rilotumab stetraxetan (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 CD D23; binds to IgE receptor), lumletuzumab (ERBB3; binds to HER3), rupartumab amadotin (binds to LYPD3), mapatumumab (binds to TRAIL-R1), margetuximab (binds to HER2), maslimomab (binds to T cell receptor), mavrilimumab (binds to GMCSF receptor α chain), matuzumab (binds to EGFR), milatuzumab (binds to CD74), minletumomab (binds to TAG-72), mirvetuximab soravtansine (binds to folate receptor alpha), mitumomab (binds to GD3 ganglioside), morolimumab (binds to rhesus factor), modotuximab (binds to EGFR extracellular domain III), mogamulizumab (binds to CCR4), monalizumab (binds to NKG2A), mosunetuzumab (binds to CD3E; MS4A1; CD20), moxetumomab pasudotox (binds to CD22), muromonab-CD3 (binds to CD3), nacolomab butafenatox (binds to C242 antigen), naptumomab estafenatox (binds to 5T4), naratuximab emtansine (binds to CD37), narutumab (binds to MST1R), natalizumab (Tysabri;binds to integrin α4), naxitamab (binds to c-Met), necitumumab (binds to EGFR), nemolizumab (binds to IL31RA), nimotuzumab (Theracim; Theraloc; binds to EGFR), nirsevimab (binds to RSVFR), nivolumab (binds to PD-1), obinutuzumab (binds to CD20), ocralizumab (binds to CD20), odulimomab (binds to LFA-1; binds to CD11a), ofatumumab (binds to CD20), olatumab (binds to PDGF-Rα), omburtamab (binds to CD276), onartuzumab (binds to human scatter factor receptor kinase), ontuxizumab (binds to TEM1), onvatilimab (binds to VSIR); Opicinumab (binding to LINGO-1), oporutozumab monatox (binding to EpCAM), oregovomab (binding to CA-125), otelixizumab (binding to CD3), otlertuzumab (binding to CD37), oxelumab (binding to OX-40), panitumumab (binding to EGFR), pancomab (binding to tumor-specific glycosylation of MUC1), patitumumab (binding to ERBB3; HER3), PDR001 (binding to PD-1), pembrolizumab (Keytruda; binding to PD-1), pemtumomab (Theragyn; binding to MUC1), pertz Ibuprofen (Omnitarg; binds to HER2 / neu), pidilizumab (binds to PD-1), pinatuzumab vedotin (binds to CD22), prosalizumab (binds to CCR2), pogalizumab (binds to TNFR superfamily member 4), polatuzumab vedotin (binds to CD79B), priligimab (binds to CD4), PRO140 (binds to CCR5), ramucirumab (Cyramza; binds to VEGFR2), ravagalimab (binds to CD40), relatolimab (binds to LAG3), linucumab (binds to platelet-derived growth factor receptor beta), Ximab (binds to CD20), rituzimab (MabThera; Rituzan; binds to CD20), lobatumumab (IGF-1 receptor; binds to CD221), racotumomab (Vaxira; binds to NGNA ganglioside), radletumab (binds to fibronectin extra domain-B), refanezumab (binds to myelin-associated glycoprotein), loredumab (binds to RHD), rovelizumab (LeukArrest; binds to CD11; binds to CD18), rozanolixizumab (binds to FCGRT), ruplizumab (Antova; binds to CD154; CD4 0L), SA237 (binding to IL-6R), sacituzumab govitecan (binding to TROP-2), samalizumab (binding to CD200), samuratamab vedotin (binding to LRRC15), sarilumab (Kevzara; binding to IL-6R), satralizumab (binding to IL6 receptor), satumomab pendetide (binding to TAG-72), serivantumab (ERBB3; binding to HER3), setrusumab (binding to SOST), SGN-CD19A (binding to CD19), SHP647 (binding to mucosal addressin cell adhesion molecule), siltuximab (Sylvant; binding to IL-6R).Soluble IL-6 (binding to IL-6R), siplizumab (binding to CD2), sirtratumab vedotin (binding to SLITRK6), sontuzumab (binding to episialin), sofituzumab vedotin (binding to CA-125), solitomab (binding to EpCAM), spartalizumab (binding to PDCD1; CD279), suresomab (binding to NCA-90; granulocyte antibody), sputumab (binding to RSVFR), tabalumab (binding to BAFF), tadocizumab (binding to integrin α; IIbβ3), talaxanthus (binding to CD123), taplitumomab paptox (binding to CD19), talectumab (binding to Notch receptors), tabolimab (binding to CD134), telisotuzumab vedotin (binding to HGFR), teneliximab (binding to CD40), tepoditamab (binding to dendritic cell-associated lectin 2), teprotumomab (binding to IGF-1 receptor; CD221), tetulomab (binding to CD37), TGN1412 (binding to CD28), tibulizumab (binding to BAFF), tigatuzumab (binding to TRAIL-R2) , timigituzumab (binds to HER2), tiragotumab (binds to TIGIT), tislelizumab (binds to PCDC1; CD279), tocilizumab (Actemra; RoActemra; binds to IL-6 receptor), tomzotuximab (EGFR; binds to HER1), toralizumab (CD154; binds to CD40L), tositumomab (Bexxar; binds to CD20), tobetumab (binds to PDGFRA), trastuzumab (Herceptin; binds to HER2 / neu), trastuzumab emtansine (Kadcyla; HER2 / n) eu), tregalizumab (binds to CD4), tremelimumab (binds to CTLA4), TRBS07 (binds to GD2 ganglioside), tucotuzumab-celmoleukin (binds to EpCAM), ublituximab (binds to MS4A1), urocupramab (CXCR4; binds to CD184), urelumab (4-1BB; binds to CD137), ustekinumab (Stellera; binds to IL-12 / 23), utomilumab (4-1BB; binds to CD137), vadastuximab-butarilin (binds to CD33), vanalimab (binds to CD40 ), vanticutumab (binds to Frizzled receptors), valisacumab (binds to VEGFR2), varlilumab (binds to CD27), batelizumab (ITGA2; binds to CD49b), vedolizumab (Entyvio; binds to integrin α4β7), veltuzumab (binds to CD20), besencumab (binds to NRP1), visilizumab (Nuvion; binds to CD3), bovalilizumab (binds to IL6R), volociximab (binds to integrin α5β1), bonlerolizumab (binds to CD134), and vopratelimab (CD278; binds to CD134).ICOS), XMAB-5574 (binding to CD19), zalutumumab (HuMax-EGFr; binding to EGFR), zanolimumab (HuMax-CD4; binding to CD4), zatuximab (binding to HER1), zenoctuzumab (ERBB3; binding to HER3), dralimumab (CD147; binding to basigin), zolbetuximab (binding to claudin-18 isoform 2), or zolimomab-allitoxin (binding to CD5), or antigen-binding portions thereof;

[0133] Other antibodies that may be secreted by cells within the device include anetuzumab (binding to mesothelin), aorutumab (binding to FGFR2), azintuximab (binding to SLAMF7), belantamab (binding to TNFRSF17), bivatuzumab (binding to CD44v6), brentuximab (binding to CD30), camidanlumab (binding to CD25), cantuzumab (binding to CanAg), clivatuzumab (binding to MUC1), cofetuzumab (binding to PTK7), and coltuximab (binding to CD 19), denintuzumab (binding to CD19), depatuximab (binding to EGFR), enapotamab (binding to AXL), enfortumab (binding to nectin-4), epratuzumab (binding to CD22), gemtuzumab (binding to CD33), glembatumumab (binding to GPNMB), hertuzumab (binding to HER2), iradatuzumab (binding to CD79B), indatuximab (binding to CD138), indutuzumab (binding to GCC), inotuzumab (binding to CD22), labetuzumab (binding to CEA-CAM4 (binding to LIV-1), laprituximab (binding to EGFR), rifastuzumab (binding to SLC34A2), loncastuzumab (binding to CD19), lorvotuzumab (binding to CD56), rosatuximab (binding to EGFR), rupartuzumab (binding to LYPD3), iratumumab (binding to CD30), milatuzumab (binding to CD74), mirvetuximab (binding to PSMA), naratuximab (binding to CD37), pinatuzumab (binding to CD22), polatuzumab (binding to CD79B), These include, but are not limited to, valpituzumab (binding to DLL3), sacituzumab (binding to TACSTD2), samtrotamab (binding to LRRC15), siltratumab (binding to SLTRK6), sofituzumab (binding to mucin 16), telisotuzumab (binding to c-Met), tisotuzumab (binding to TF), trastuzumab (binding to ERBB2), vadastuximab (binding to CD33), bundustuzumab (binding to STEAP1), or borsetuzumab (binding to CD70), or antigen-binding portions thereof.

[0134] In some embodiments, the cells in the cell chamber device secrete an antibody that specifically binds to α4β7, or an antigen-binding portion thereof. In certain embodiments, the cells secrete vedolizumab, or an antigen-binding portion thereof. In some embodiments, the cells in the device are mammalian host cells engineered to stably express an anti-α4β7 antibody, such as vedolizumab, or a binding molecule containing the antigen-binding region of vedolizumab. Vedolizumab is also known by its trade name ENTYVIO® (Millennium Pharmaceuticals, Inc.). Vedolizumab is a humanized monoclonal antibody that specifically binds to α4β7 integrin, e.g., the α4β7 complex, blocks the interaction of α4β7 integrin with mucosal addressin cell adhesion molecule-1 (MAdCAM-1) and fibronectin, and inhibits transendothelial migration of lymphocytes, e.g., CD4, CD8, and memory T lymphocytes, into inflamed gastrointestinal parenchymal tissue. Vedolizumab does not bind to or inhibit the function of α4β1 and αEβ7 integrins, and does not antagonize the interaction of α4 integrin with vascular cell adhesion molecule-1 (VCAM-1).

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

[0136] In one embodiment, the cells in the cell chamber device secrete an antibody, or an antigen-binding portion thereof, comprising the heavy chain variable region of SEQ ID NO: 1 and / or the light chain variable region of SEQ ID NO: 5. In one embodiment, the cells in the cell chamber device secrete an antibody, or an antigen-binding portion thereof, comprising the heavy chain CDR1 of SEQ ID NO: 2, the heavy chain CDR2 of SEQ ID NO: 3, and the heavy chain CDR3 of SEQ ID NO: 4, and / or the light chain CDR1 of SEQ ID NO: 6, the light chain CDR2 of SEQ ID NO: 7, and the light chain CDR3 of SEQ ID NO: 8. In one embodiment, the cells in the cell chamber device secrete an antibody, or an antigen-binding portion thereof, comprising 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 the sequence of vedolizumab are also described in U.S. Patent Publication Nos. 2014 / 0341885 and 2014 / 0377251, each of which is expressly incorporated by reference in its entirety. The cells in the devices herein can be engineered to secrete antibodies comprising the binding regions, eg, CDRs or variable regions, set forth above and in the enclosed sequence listing.

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

[0138] In one embodiment, cells in the cell chamber device secrete an antibody, or antigen-binding portion thereof, that specifically binds to tumor necrosis factor alpha (TNFα). In one embodiment, cells in the cell chamber device secrete an antibody, or antigen-binding portion thereof, comprising the heavy chain variable region of SEQ ID NO: 22 and / or the light chain variable region of SEQ ID NO: 23. In one embodiment, cells in the cell chamber device secrete an antibody, or antigen-binding portion thereof, comprising the heavy chain CDR1 of SEQ ID NO: 24, the heavy chain CDR2 of SEQ ID NO: 25, and the heavy chain CDR3 of SEQ ID NO: 26, and / or the light chain CDR1 of SEQ ID NO: 27, the light chain CDR2 of SEQ ID NO: 28, and the light chain CDR3 of SEQ ID NO: 29. In one embodiment, cells in the cell chamber device secrete an antibody, or antigen-binding portion thereof, comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 30 and / or a light chain comprising the amino acid sequence of SEQ ID NO: 31. Cells in the devices herein can be engineered to secrete antibodies comprising the binding regions, e.g., CDRs or variable regions, set forth above and in the enclosed sequence listing.

[0139] In one embodiment, cells in the cell chamber device secrete an antibody, or antigen-binding portion thereof, that specifically binds to interleukin-12 (IL-12). In one embodiment, cells in the cell chamber device secrete an antibody, or antigen-binding portion thereof, comprising the heavy chain variable region of SEQ ID NO: 32 and / or the light chain variable region of SEQ ID NO: 33. In one embodiment, cells in the cell chamber device secrete an antibody, or antigen-binding portion thereof, comprising the heavy chain CDR1 of SEQ ID NO: 34, the heavy chain CDR2 of SEQ ID NO: 35, and the heavy chain CDR3 of SEQ ID NO: 36, and / or the light chain CDR1 of SEQ ID NO: 37, the light chain CDR2 of SEQ ID NO: 38, and the light chain CDR3 of SEQ ID NO: 39. In one embodiment, cells in the cell chamber device secrete an antibody, or antigen-binding portion thereof, comprising 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. Cells in the devices herein can be engineered to secrete antibodies comprising the binding regions, e.g., CDRs or variable regions, set forth above and in the enclosed sequence listing.

[0140] In some embodiments, the cell chamber device comprises cells comprising one or more nucleic acid molecules encoding an antibody, or antigen-binding portion thereof, e.g., an antibody described above, or an antigen-binding portion thereof, operably linked to a promoter and / or other elements necessary for gene expression. In one embodiment, the cell chamber device comprises a recombinant host cell comprising one or more expression vector(s) comprising one or more nucleic acid(s) encoding an antibody heavy chain and / or antibody light chain, or portion thereof.

[0141] For example, in some embodiments, the cell chamber device may be configured to bind to 3F8 (binds to GD2 ganglioside), abciximab (ReoPro; binds to CD41), abituzumab (binds to CD51), alemtuzumab (Lemtrada, Campath; binds to CD52), abrilumab (binds to integrin α4β7), adalimumab (Humira; binds to TNF-α), adecatumumab (binds to EpCAM), alacizumab pegol (binds to VEGFR2), alemtuzumab (binds to EpCAM), alacizumab pegol (binds to VEGFR2), alemtuzumab (binds to EpCAM), alc ...lemtuzumab (binds to EpCAM), alemtuzumab (binds to EpCAM), alemtuzumab (binds to EpCAM), alemtuzumab (binds to EpCAM), alemtuzumab (binds to EpCAM), alemtuzumab (binds to EpCAM), alemtuzumab (binds to EpCAM), alemtuz Mabs (Lemtrada, Campath; binds to CD52), altumomab pentetate (Hybri-ceaker; binds to CEA), amatuximab (binds to mesothelin), anatumomab mafenatox (binds to TAG-72), anetumab ravtansine (binds to MSLN), anifrolumab (binds to interferon α / β receptor), apolizumab (binds to HLA-DR), apolizumab ixadotin (binds to FGFR2), arcitumomab (binds to CEA) combination), acelizumab (binding to L-selectin or CD62L), atezolizumab (Tecentriq; binding to PD-L1), atrolimumab (binding to rhesus factor), avelumab (Bavencio; binding to PD-L1), abicixizumab (binding to DLL4; VEGFA), azintuximab vedotin (binding to CD319), basiliximab (Simulect; binding to CD25), bavituximab (binding to phosphatidylserine), BCD-100 (P D-1), vectamumomab (LymphoScan; binds to CD22), belantamab mafodotin (binds to BCMA), belimumab (Benlysta; binds to BAFF), bemarituzumab (binds to FGFR2), benralizumab (Fasenra; binds to CD125), velsanlimab (binds to ICAM-1), besilesomab (Scintimun; binds to CEA-related antigen), bimagrumab (binds to ACVR2B), bivatuzumab mertansine (CD44 v6), bleselumab (binds to CD40), blinatumomab (Blincyto; binds to CD19), brosozumab (binds to SOST), brentuximab vedotin (Adcentris; binds to CD30), brontuzumab (binds to Notch1), brodalumab (Siliq;IL-17 receptor), cabilalizumab (CSF1R), camidanlumab tesirin (CD25), camrelizumab (PD-1), carotuximab (endoglin), catumaxomab (Removab; EpCAM / CD3), cantuzumab ravtansine (MUC1), caplacizumab (Cablivi; VWF), cedelizumab (CD4), semipilimab (Libtayo; PCDC1), and ceto Relimab (binds to PD-1), certolizumab (binds to TNF-α), sergituzumab amnalikine (binds to CEA), cetuximab (Erbitux; binds to EGFR), sibisatamab (binds to CEACAM5), cirumutuzumab (binds to ROR1), cixutumumab (binds to IGF-1 receptor; CD221), clenoliximab (binds to CD4), clivatuzumab tetraxetan (hPAM4-Cide; binds to MUC1), codrituzumab (binds to glypican 3), coltuximab ravtansine (binds to CD19), conatumumab (binds to TRAIL-R2), crizanlizumab (binds to selectin P), clotidumab (binds to GCGR), dacetuzumab (binds to CD40), daclizumab (Zenapax; binds to CD25), dalotuzumab (IGF-1 receptor; binds to CD221), dapirolizumab pegol (binds to CD154; binds to CD40L), daratumumab (Darzalex; binds to CD38), dem Tucizumab (binding to DLL4), denintuzumab mafodotin (binding to CD19), depatuxizumab mafodotin (binding to EGFR), drozizumab (binding to DR5), DS-8201 (binding to HER2), deligotuzumab (ERBB3; binding to HER3), dinutuximab (Unituxin; binding to GD2 ganglioside), dupilumab (binding to IL-4Rα), durvalumab (Imfinzi; binding to PD-L1), duvoltuximab (CD19;CD3E), ecloneximab (binding to GD3 ganglioside), edrecolomab (binding to EpCAM), elezanumab (binding to RGMA), elgemuzumab (binding to ERBB3, HER3), elotuzumab (binding to SLAMF7), emactuzumab (binding to CSF1R), enapotamab vedotin (binding to AXL), enavatuzumab (binding to TWEAK receptor) binds), enlimonomab pegol (ICAM-1; binds to CD54), enoblituumab (binds to CD276), enoticumab (binds to DLL4), epratuzumab (binds to CD22), erlizumab (ITGB2; binds to CD18), ertumakizumab (Rexomun; HER2 / neu; binds to CD3), etaracizumab (Abergin; integrin α; vβ3), etigilimab (binding to TIGIT), etrolizumab (binding to integrin β7), exbivirumab (binding to hepatitis B surface antigen), fanolesomab (binding to NeutroSpec; binding to CD15), faralimomab (binding to interferon receptor), farletuzumab (binding to folate receptor 1), FBTA05 (Lymphomun; binding to CD20), fugatipotuzumab (binding to MUC1), fimatuzumab (binding to ephrin receptor A3), fizitumumab (binding to IGF-1 receptor; CD221), Lotetuzumab (binding to IL3 receptor), foralumab (binding to CD3 epsilon), futuximab (binding to EGFR), galiximab (binding to CD80), gancotamab (binding to HER2 / neu), ganitumab (binding to IGF-1 receptor; CD221), gavilimomab (binding to CD147; basigin), gemtuzumab ozogamicin (Mylotarg; binding to CD33), glenbatumumab vedotin (binding to GPNMB), golimumab (Simponi; binding to TNF-α), gomiliximab (CD23; binding to IgE receptor), ianalumab (BAF F-R), ibalizumab (Trogarzo; binds to CD4), IBI308 (binds to PD-1), ibritumomab tiuxetan (binds to CD20), icrucumab (binds to VEGFR-1), ifavotuzumab (binds to EPHA3), igovomab (Indimacis-125; binds to CA-125), IMAB362 (binds to CLDN18.2), imaprelimab (binds to MCAM), inlacumab (binds to selectin P), indatuximab ravtansine (binds to SDC1), iradatuzumab vedotin (binds to CD97B), imgatuzumab (binds to EGFR), indusatumab vedotin (binds to GUCY2C), inebilizumab (binds to CD19), infliximab (Remicade; binds to TNF-α), intetumumab (binds to CD51), inolimomab (binds to CD25), inotuzumab ozogamicin (Besponsa; binds to CD22), ipilimumab (Yervoy; binds to CD152), iomab-B (binds to CD45), iratumumab (binds to CD30), isatuximab (binds to CD38), iscalimab (binds to CD40), istiratumab (binds to IGF1R;CD221), Itolizumab (Alzumab; binds to CD6), Keliximab (binds to CD4), Laprituximab Emtansine (binds to EGFR), Labetuzumab (CEA-Cide; binds to CEA), Rifastuzumab Vedotin (binds to sodium phosphate cotransporter), Remaresomab (NCA-90; binds to granulocyte antigen), Lembervimab (binds to hepatitis B surface antigen), Leronlimab (binds to CCR5), Lexatumumab (binds to TRAIL-R2), Ribivirumab (binds to hepatitis B surface antigen), Loncastuximab Tecilin (binds to CD19) Binds to EGFR; ERBB1; HER1), rirotuzumab satetraxetan (binds to CD37), lintuzumab (binds to CD33), lirilumab (binds to KIR2D), lorvotuzumab mertansine (binds to CD56), lucatumumab (binds to CD40), lulizumab pegol (binds to CD28), rumiliximab (binds to CD23; binds to IgE receptor), ramletuzumab (binds to ERBB3; HER3), rupartumab amadotin (binds to LYPD3), mapatumumab (binds to TRAIL-R1), margetuximab Mab (binding to HER2), maslimomab (binding to T cell receptor), mavrilimumab (binding to GMCSF receptor α chain), matuzumab (binding to EGFR), milatuzumab (binding to CD74), minletumomab (binding to TAG-72), mirvetuximab soravtansine (binding to folate receptor α), mitumomab (binding to GD3 ganglioside), morolimumab (binding to rhesus factor), modotuximab (binding to EGFR extracellular domain III), mogamulizumab (binding to CCR4), monalizumab (binding to NKG2A), mosunetuzumab (binding to CD3E; MS4 A1; binds to CD20), moxetumomab pasudotox (binds to CD22), muromonab-CD3 (binds to CD3), nacolomab butafenatox (binds to C242 antigen), natumomab estafenatox (binds to 5T4), naratuximab emtansine (binds to CD37), narunatumab (binds to MST1R), natalizumab (Tysabri; binds to integrin α4), naxitamab (binds to c-Met), necitumumab (binds to EGFR), nemolizumab (binds to IL31RA), nimotuzumab (Theracim; Theraloc;EGFR), nirsevimab (binding to RSVFR), nivolumab (binding to PD-1), obinutuzumab (binding to CD20), ocralizumab (binding to CD20), ocrelizumab (binding to CD20), odulimomab (binding to LFA-1; CD11a), ofatumumab (binding to CD20), olaratumab (binding to PDGF-Rα), omburtamab (binding to CD276), onartuzumab (binding to human scatter factor receptor kinase), ontuximab (binding to TEM1) ), onvatilimab (binding to VSIR), opicinumab (binding to LINGO-1), oporutozumab monatox (binding to EpCAM), oregovomab (binding to CA-125), otelixizumab (binding to CD3), otlertuzumab (binding to CD37), oxelumab (binding to OX-40), panitumumab (binding to EGFR), pancomab (binding to tumor-specific glycosylation of MUC1), patitumumab (binding to ERBB3; HER3), PDR001 (binding to PD-1); Pembrolizumab (Keytruda; binds to PD-1), pemtumomab (Theragyn; binds to MUC1), pertuzumab (Omnitarg; binds to HER2 / neu), pidilizumab (binds to PD-1), pinatuzumab vedotin (binds to CD22), prosalizumab (binds to CCR2), pogalizumab (binds to TNFR superfamily member 4), polatuzumab vedotin (binds to CD79B), priligimab (binds to CD4), PRO140 (binds to CCR5), ramucirumab (Cyramza; VEGFR2 (binding to CD40), ravagalimab (binding to CD40), raletimab (binding to LAG3), linucumab (binding to platelet-derived growth factor receptor beta), rituximab (binding to CD20), rituzimab (MabThera; Rituzan; binding to CD20), lobatumumab (IGF-1 receptor; binding to CD221), racotumomab (Vaxira; binding to NGNA ganglioside), radletumab (binding to fibronectin extradomain-B), refanezumab (binding to myelin-associated glycoprotein), loredumab (binding to RHD), Belizumab (LeukArrest; binds to CD11; CD18), rozanolixizumab (binds to FCGRT), ruplizumab (Antova; binds to CD154; CD40L), SA237 (binds to IL-6R), sacituzumab govitecan (binds to TROP-2), samalizumab (binds to CD200), samuratamab vedotin (binds to LRRC15), sarilumab (Kevzara; binds to IL-6R), satralizumab (binds to IL6 receptor), satumomab pendetide (binds to TAG-72), seribantumab (ERBB3; binds to ERBB3) HER3), setrusumab (binds to SOST), SGN-CD19A (binds to CD19), SHP647 (binds to mucosal addressin cell adhesion molecule), silmutuzumab (Sylvant; binds to soluble IL-6, IL-6R), siplizumab (binds to CD2), siltratumab vedotin (binds to SLITRK6), sontuzumab (binds to episialin), sofituzumab vedotin (binds to CA-125), solitomab (binds to EpCAM), spartalizumab (PDCD1; binds to CD279), suletomab (NCA-90; binds to PDCD1).granulocyte antibodies), sputabumab (binding to RSVFR), tabalumab (binding to BAFF), tadocizumab (binding to integrin α; IIbβ3), talaxanthus (binding to CD123), taplitumomab paptox (binding to CD19), tarexuzumab (binding to Notch receptor), tabolimab (binding to CD134), telisotuzumab vedotin (binding to HGFR), teneliximab (binding to CD40), tepoditamab (binding to dendritic cell-associated lectin 2), teprotumomab (binding to IGF-1 receptor; CD221), tetulomab (binding to CD37), TGN1412 (binding to CD28), tiburizumab (binding to BAFF), tigatuzumab (binding to TRAIL-R2), Timigtuzumab (binds to HER2), tiragotumab (binds to TIGIT), tislelizumab (binds to PCDC1; CD279), tocilizumab (Actemra; RoActemra; binds to IL-6 receptor), tomzotuximab (EGFR; binds to HER1), toralizumab (CD154; binds to CD40L), tositumomab (Bexxar; binds to CD20), tobetumab (binds to PDGFRA), trastuzumab (Herceptin; binds to HER2 / neu), trastuzumab emtansine (Kadcyla; HER2 / n) eu), tregalizumab (binds to CD4), tremelimumab (binds to CTLA4), TRBS07 (binds to GD2 ganglioside), tucotuzumab-celmoleukin (binds to EpCAM), ublituximab (binds to MS4A1), urocuplumab (binds to CXCR4; CD184), urelumab (4-1BB; binds to CD137), ustekinumab (Stellera; binds to IL-12 / 23), utomilumab (4-1BB; binds to CD137), vadastuximab-butarilin (binds to CD33), vanalimab (binds to CD40 ), vanticutumab (binds to Frizzled receptors), valisacumab (binds to VEGFR2), varlilumab (binds to CD27), batelizumab (ITGA2; binds to CD49b), vedolizumab (Entyvio; binds to integrin α4β7), veltuzumab (binds to CD20), besencumab (binds to NRP1), visilizumab (Nuvion; binds to CD3), bovalilizumab (binds to IL6R), volociximab (binds to integrin α5β1), bonlerolizumab (binds to CD134), vopratelimab (CD278;The chamber may contain cells containing one or more nucleic acid molecules encoding one or more of the following: ICOS (binding to ICOS), XMAB-5574 (binding to CD19), zalutumumab (HuMax-EGFr; binding to EGFR), zanolimumab (HuMax-CD4; binding to CD4), zatuximab (binding to HER1), xenoctuzumab (ERBB3; binding to HER3), dralimumab (CD147; binding to basigin), zolbetuximab (binding to claudin-18 isoform 2), or zolimomab alitox (binding to CD5), or antigen-binding portions thereof. Preferably, the nucleic acid molecule is operably linked to a promoter and / or other regulatory elements necessary for expression and secretion of the protein encoded by the nucleic acid by the cells present in the chamber.

[0142] In some embodiments, cells in a cell chamber device disclosed herein comprise one or more nucleic acids encoding an antibody that specifically binds to α4β7, or an antigen-binding portion thereof. In certain embodiments, cells comprise one or more nucleic acids encoding vedolizumab or an antigen-binding portion of vedolizumab. The nucleic acid sequence encoding the light chain variable region is set forth in SEQ ID NO: 11. 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 vedolizumab is set forth as SEQ ID NO: 13, and the full-length nucleic acid sequence encoding the heavy chain of vedolizumab is set forth as SEQ ID NO: 14. Alternative nucleic acid sequences encoding vedolizumab are set forth in U.S. Patent Publication No. 2010 / 0297699, which is incorporated herein in its entirety. Thus, in certain embodiments, cells in a cell chamber device can comprise the nucleic acid sequence set forth in SEQ ID NO: 11 and / or the nucleic acid sequence set forth in SEQ ID NO: 12. In some embodiments, cells in a cell chamber device comprise 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, cells in a cell chamber device disclosed herein express one or more nucleic acids encoding an antibody that specifically binds to α4β7, or an antigen-binding portion thereof. In certain embodiments, the cells express one or more nucleic acids encoding vedolizumab or an antigen-binding portion of vedolizumab. In certain embodiments, cells in a cell chamber device express the nucleic acid sequence set forth in SEQ ID NO: 11 and / or the nucleic acid sequence set forth in SEQ ID NO: 12. In some embodiments, cells in a cell chamber device express a nucleic acid comprising the nucleic acid sequence of SEQ ID NO: 13 and the nucleic acid sequence of SEQ ID NO: 14.

[0144] In some embodiments, the cells in the cell chamber device express a nucleic acid comprising the nucleic acid sequence of an immature humanized anti-α4β7 immunoglobulin chain containing a signal peptide (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 nucleic acid comprising the nucleic acid sequence of a mature humanized anti-α4β7 immunoglobulin chain without a signal peptide (e.g., nucleotides 77-1429 of SEQ ID NO: 15, nucleotides 79-735 of SEQ ID NO: 16, nucleotides 76-1428 of SEQ ID NO: 17, nucleotides 78-734 of SEQ ID NO: 18, nucleotides 58-714 of SEQ ID NO: 19, or nucleotides 58-1410 of SEQ ID NO: 20).

[0145] In one embodiment, the cells in the cell chamber device comprise a recombinant expression vector, the recombinant expression vector comprising a first nucleic acid encoding an immunoglobulin heavy chain and a second nucleic acid encoding an immunoglobulin light chain, the first nucleic acid comprising nucleotides 77-1429 of SEQ ID NO: 15 and the second nucleic acid comprising nucleotides 79-735 of SEQ ID NO: 16.

[0146] In one embodiment, the cells in the cell chamber device comprise a recombinant expression vector, the recombinant expression vector comprising a first nucleic acid encoding an immunoglobulin heavy chain and a second nucleic acid encoding an immunoglobulin light chain, the first nucleic acid comprising nucleotides 76-1428 of SEQ ID NO: 17 and the second nucleic acid comprising nucleotides 78-734 of SEQ ID NO: 18.

[0147] In one embodiment, the cells in the cell chamber device comprise a recombinant expression vector, the recombinant expression vector comprising a first nucleic acid encoding an immunoglobulin heavy chain and a second nucleic acid encoding an immunoglobulin light chain, the first nucleic acid comprising nucleotides 58-1410 of SEQ ID NO: 20 and the second nucleic acid comprising nucleotides 58-714 of SEQ ID NO: 19.

[0148] Additional examples of nucleic acid sequences that can be expressed by cells within a cell chamber device are also described in WO2008 / 115504, which is incorporated herein by reference in its entirety.

[0149] In an exemplary embodiment, the cell chamber device contains ARPE-19 cells that secrete an antibody, or antigen-binding portion thereof, comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 5. In some embodiments, the cell chamber device contains ARPE-19 cells that secrete an antibody, or antigen-binding portion thereof, comprising a heavy chain variable region comprising CDR1 of SEQ ID NO: 2, CDR2 of SEQ ID NO: 3, and CDR3 of SEQ ID NO: 4, and a light chain variable region comprising CDR1 of SEQ ID NO: 6, CDR2 of SEQ ID NO: 7, and CDR3 of SEQ ID NO: 8. In some embodiments, the cell chamber device contains ARPE-19 cells that secrete an antibody, or antigen-binding portion thereof, 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 cell chamber device contains ARPE-19 cells that contain a nucleic acid 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 contains ARPE-19 cells that contain 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 contains ARPE-19 cells that secrete vedolizumab.

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

[0151] In some embodiments, cells in a cell chamber device disclosed herein express one or more nucleic acids encoding an antibody, or antigen-binding portion thereof, that specifically binds to IL-12. In certain embodiments, cells in a cell chamber device express the nucleic acid sequence set forth in SEQ ID NO: 46 and / or the nucleic acid sequence set forth in SEQ ID NO: 47. In some embodiments, cells in a cell chamber device express a nucleic acid comprising the nucleic acid sequence of SEQ ID NO: 48 and / or the nucleic acid sequence of SEQ ID NO: 49. In one embodiment, cells in a cell chamber device comprise a recombinant expression vector, wherein the recombinant expression vector comprises a first nucleic acid encoding an immunoglobulin heavy chain and a second nucleic acid encoding an immunoglobulin light chain, wherein 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 can secrete peptide therapeutics for gastrointestinal use, such as for the treatment of short bowel syndrome. Examples of peptide therapeutics useful for treating gastrointestinal disorders are also described in US 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., SEQ ID NO: 8). Each of the foregoing patents and patent applications is incorporated herein by reference in its entirety.

[0153] The amino acid sequence of an exemplary peptide therapeutic for gastrointestinal use is provided herein as SEQ ID NO: 21. In certain embodiments, cells in the cell chamber secrete a peptide having the amino acid sequence of SEQ ID NO: 21. In some embodiments, cells in the cell chamber devices disclosed herein comprise a nucleic acid encoding a peptide having the amino acid sequence of SEQ ID NO: 21.

[0154] In certain embodiments, the cell chamber device contains ARPE-19 cells that secrete a peptide having the amino acid sequence of SEQ ID NO: 21. In some embodiments, the cell chamber device contains ARPE-19 cells that contain a nucleic acid encoding a peptide having the amino acid sequence of SEQ ID NO: 21.

[0155] Cells can be engineered to produce biomolecules, such as those described herein, by inserting an expression construct encoding the biomolecule into the cells using standard techniques. For example, a vector containing a polynucleotide encoding a polypeptide of interest can be inserted into cells to generate a cell line that produces the polypeptide of interest. The term "vector," as used herein, is intended to refer to a vehicle, e.g., a nucleic acid molecule, that can deliver genetic material to a cell, where it can then be replicated and / or integrated into the cellular genome and expressed. One type of vector is a plasmid, which refers to a circular double-stranded DNA into which additional DNA segments can be ligated. Other common vectors include phage vectors and viral vectors. Certain vectors are capable of autonomous replication within a host cell into which they are introduced (e.g., episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) can be integrated into the genome of the host cell upon introduction into the host cell, thereby replicating along with the host genome. Furthermore, certain vectors are capable of directing the expression of genes to which they are operably linked. Such vectors are referred to herein as "recombinant expression vectors," or simply "expression vectors," or "expression constructs." A variety of vectors suitable for the recombinant expression of therapeutic proteins, e.g., therapeutic antibodies, are publicly available and well known to those skilled in the art. Vector components generally include, but are not limited to, one or more of a signal sequence, an origin of replication, one or more marker genes, an enhancer element, a promoter, and a transcription termination sequence. Optional signal sequences, origins of replication, marker genes, enhancer elements, and transcription terminator sequences that can be used are known in the art and are described in further detail, for example, in U.S. Pat. No. 7,053,202. In certain cases, the promoter element driving expression of the biomolecule is a constitutive promoter element to ensure long-term expression of the biomolecule in the cells of the device.Expression of a biomolecule can also be increased by various other methods known in the art, for example, by increasing the copy number of the nucleic acid expressing the biomolecule, by removing premature termination or splicing signals within the nucleic acid expressing the biomolecule, or by selecting a promoter for the site of genomic integration that allows for increased expression of the biomolecule. Another example of increasing expression levels is targeted integration into transcription hotspots, for example, by homologous recombination, determined empirically by genome sequencing or computationally by expression probability algorithms.

[0156] IV. Three-dimensional cell culture Also provided herein is a cell chamber device comprising a nanofibrous (e.g., electrospun) polymer capable of accommodating three-dimensional cell culture. Thus, in some embodiments, the device can contain a tissue or a portion thereof (e.g., a tissue explant) having a three-dimensional structure, or a cluster of cells (e.g., an organoid or spheroid). A cell chamber device suitable for three-dimensional cell culture can be useful, for example, in a therapeutic method in which secretion of a biomolecule endogenously produced by a cell or tissue is desired. Cells or tissues can be grown in three-dimensional cell culture (i.e., 3D cell culture) according to methods known in the art (see, for example, Edmondson, et al. (2014). Assay and drug development technologies, 12(4), 207-218, incorporated herein by reference), and then added to the cell chamber device. Cells or tissues having a three-dimensional structure can be obtained, for example, from tissue explants, biopsies, or harvested from living donors, cell cultures, or autopsies using art-recognized techniques.

[0157] In some embodiments, the cell chamber device includes a three-dimensional cell culture comprising a tissue, or a portion thereof. Examples of tissues that can be included in the devices herein include liver tissue, pancreatic tissue, intestinal tissue, or kidney tissue. The tissue can optionally be a tissue explant (e.g., a piece or pieces of tissue or organ removed from an animal). In one embodiment, the cell chamber includes liver tissue (e.g., a liver tissue explant). In one embodiment, the cell chamber includes pancreatic tissue (e.g., a pancreatic tissue explant, such as an islet of Langerhans tissue explant). In one embodiment, the cell chamber includes kidney tissue (e.g., a kidney tissue explant). In some embodiments, the cell chamber includes reproductive tissue (e.g., ovarian or testicular tissue). In some embodiments, the tissue is human tissue, such as a human kidney tissue explant, a human liver tissue explant, or human pancreatic tissue. In other embodiments, the tissue is human reproductive tissue, such as a human ovarian or testicular explant.

[0158] Alternatively, the cell chamber device may contain a cluster of cells with a three-dimensional structure. For example, in some embodiments, the three-dimensional cell culture may contain organoids or spheroids. In some embodiments, the cluster of cells (e.g., organoids or spheroids) comprises hepatocytes (e.g., human hepatocytes), kidney cells, or islet cells of Langerhans. The cluster of cells (e.g., organoids or spheroids) may be organized around a structure to maintain the three-dimensional structure of the cell culture. For example, in some embodiments, the cell chamber comprises an organized group of cells organized around a structure such as a tube or sinusoid.

[0159] A cell chamber device for use with three-dimensional cell culture may include a multilayer scaffold comprising a nanofibrous, e.g., electrospun, polymer, as described herein (see, e.g., Section II). Optionally, the scaffold layers can be tailored to promote angiogenesis toward tissue or cells within the chamber.

[0160] In alternative embodiments, the cell chamber device can include a single nanofibrous, e.g., electrospun, polymer layer. Monolayer scaffolds can be formed from a variety of polymers (e.g., nanofibrous polymers), such as polyester, polyethylene terephthalate (PET, also known as Dacron), polybutylene terephthalate (PBT), or polyurethane (PU). For example, in some embodiments, the monolayer scaffold can include nanofibrous polyester, nanofibrous polyethylene terephthalate (nPET), nanofibrous polybutylene terephthalate (nPBT), and / or nanofibrous polyurethane (nPU). In one embodiment, the monolayer scaffold includes nPET. In another embodiment, the monolayer scaffold includes nPBT. In one embodiment, the monolayer scaffold includes nPET-nPBT. In a further embodiment, the monolayer scaffold includes nPU. Monolayer cell chamber devices can be fabricated essentially as described for the multilayer devices provided herein, using a polymer sheet including a single, e.g., homogeneous, layer of nanofibrous polymer. In some embodiments, a single layer device can allow for greater contact between cells within the chamber and the environment outside the chamber compared to a multi-layer scaffold device.

[0161] In some embodiments, the nanofibrous polymer scaffold can be loaded with an anti-inflammatory agent. For example, the nanofibrous polymer scaffold can be loaded with tacrolimus, pirfenidone, and / or roflumilast. Without wishing to be bound by theory, the inclusion of an anti-inflammatory agent may help preserve the three-dimensional structure of cells growing within the chamber by preventing the infiltration of cells (e.g., immune cells or inflammatory cells) from outside the chamber. The anti-inflammatory agent can also reduce a subject's immune response to the device after implantation. In some embodiments, the cell chamber device includes a single-layer scaffold comprising a nanofibrous polymer, wherein the scaffold comprises an anti-inflammatory agent (e.g., tacrolimus, pirfenidone, and / or roflumilast). In other embodiments, the cell chamber device includes a multi-layer scaffold comprising a nanofibrous polymer, wherein the scaffold comprises an anti-inflammatory agent (e.g., tacrolimus, pirfenidone, and / or roflumilast).

[0162] The tissue graft or cells having a three-dimensional structure can optionally secrete a biomolecule. In some embodiments, the biomolecule is a biomolecule endogenously produced by the tissue or cell. In alternative embodiments, the cells loaded into the cell chamber device of the present disclosure can be genetically engineered cells, e.g., recombinant cells, engineered to produce, e.g., secrete, a biomolecule of interest. Biomolecules that can be secreted by cells in the device include, but are not limited to, polypeptides, polysaccharides, and polynucleotides, as well as organic molecules such as 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, nucleotides, deoxyribonucleic acid (DNA), or ribonucleic acid (RNA). Other secreted biomolecules can include those assembled, packaged, and secreted as exosomes, lipid polymers, or viral particles. In some embodiments, the tissue graft or cells secrete a therapeutic biomolecule, as described herein.

[0163] In some embodiments, tissues or cells within the device (e.g., cells having a three-dimensional structure) secrete one or more proteins or peptides, e.g., one or more therapeutic proteins or peptides. For example, the tissues or cells within the device can secrete one or more therapeutic proteins, such as an antibody or antigen-binding fragment thereof, a growth factor, a hormone (e.g., insulin), a cytokine, a clotting factor (e.g., Factor VIII or Factor IX, or a variant thereof, e.g., Recombinate, Kogenate, Refacto, Advate, Alprolix, BeneFIX, Rixubis, Ixinity, Idelvion, etc.), or a combination thereof.

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

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

[0166] In another embodiment, the cell chamber device includes liver tissue or hepatocytes having a three-dimensional structure, and the liver tissue or hepatocytes secrete albumin, transferrin, plasminogen, fibrinogen, alpha-fetoprotein, and / or clotting factors. In another embodiment, the liver tissue or hepatocytes can secrete digestive enzymes and products, such as bile, alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), and / or gamma-glutamyl transpeptidase (GGT).

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

[0168] In some embodiments, the tissues or cells within the device can secrete one protein or peptide, e.g., one endogenous or recombinant protein or peptide. In other embodiments, the cells within the device can secrete two or more proteins, e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10 or more endogenous or recombinant proteins. In some embodiments, the device can be loaded with a single tissue or cell cluster having a three-dimensional structure, e.g., a single tissue or cell cluster, that secretes multiple proteins or peptides.

[0169] V. Methods of Administration and Treatment Also provided herein are methods for delivering biomolecules to a subject, comprising administering to the subject a cell chamber device disclosed herein, wherein the cell chamber of the device contains cells that secrete the biomolecule. The biomolecule may be, for example, a recombinant peptide or recombinant protein, including those described above. In certain cases, this method can be used to deliver, for example, an antibody, such as an anti-α4β7 antibody (e.g., vedolizumab), or an antigen-binding portion thereof, to a subject. The dosage of the biomolecule delivered by the cell chamber device containing the biomolecule-producing cells can be controlled by varying the dimensions (length, diameter, volume) of the cell chamber device, adjusting the number of cells within the device via geometry, adjusting the expression level of the biomolecule by the cells (e.g., by altering copy number, selecting a promoter, etc.), and / or adjusting the number of devices delivered to the subject (e.g., 1 to 10 devices per patient). Thus, a subject may be administered one or more of the cell chamber devices described herein. In some embodiments, a subject is administered one cell chamber device. In other embodiments, the subject is administered 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 at multiple sites within the body.

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

[0171] In some embodiments, the device is administered in a concentration of about 1×10 4 cells ~ approx. 1 x 10 12 cells (e.g., approximately 1 x 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 cells ~ approx. 1 x 10 9 cells, or approximately 1 x 109 cells ~ approx. 1 x 10 12 In some embodiments, the device is loaded with, and therefore capable of containing, about 1 x 10 cells. 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 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 x 10 6 cells ~ approx. 1 x 10 7 cells, approximately 6 x 10 6 cells ~ approx. 9 x 10 6 cells, approximately 7 x 10 6 cells ~ approx. 9 x 10 6 cells, or approximately 8 x 10 6 cells ~ approx. 9 x 10 6 For example, in certain embodiments, the device contains approximately 8.5 x 10 6 In some embodiments, the device contains about 1 x 10 cells. 7 cells ~ approx. 1 x 10 8 In some embodiments, the device contains about 1 x 10 cells. 8 cells ~ approx. 1 x 10 9 In some embodiments, the device contains about 1 x 10 cells. 9 cells ~ approx. 1 x 10 10 In some embodiments, the device contains about 1 x 10 cells. 10 cells ~ approx. 1 x 10 11 In some embodiments, the device contains about 1 x 10 cells. 11 cells ~ approx. 1 x 10 12 In an exemplary embodiment, the device contains about 1 x 10 cells. 4 cells, 1 x 10 5 cells, 1 x 10 6 cells, 1 x 10 7 cells, 1 x 10 8cells, 1 x 10 9 cells, 1 x 10 10 cells, 1 x 10 11 cells, or 1 x 10 12 In some embodiments, the device contains about 1 x 10 cells prior to implantation into the subject. 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 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 cells / cm 2 , 1×10 5 cells / cm 2 ~Approx. 1×10 6 cells / cm 2 , about 1×10 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 cells / cm 2 , about 5×10 5 cells / cm 2 ~Approx. 7×10 5 cells / cm 2 , about 6×10 5 cells / cm 2 ~Approx. 7×10 5 cells / cm 2 , about 6×10 5 cells / cm 2 ~Approx. 6.5×10 5 cells / cm2 , or 6.25 × 10 5 cells / cm 2 Includes.

[0173] In other embodiments, the device may contain approximately 1×10 cells after implantation in a subject (e.g., after a period of time sufficient for the cells in the chamber to proliferate). 4 cells ~ approx. 1 x 10 12 cells (e.g., approximately 1 x 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 cells ~ approx. 1 x 10 9 cells, or approximately 1 x 10 9 cells ~ approx. 1 x 10 12 In some embodiments, the device has a capacity to accommodate about 1 x 10 cells. 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 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 x 10 6 cells ~ approx. 1 x 10 7 cells, approximately 6 x 10 6 cells ~ approx. 9 x 10 6 cells, approximately 7 x 10 6 cells ~ approx. 9 x 10 6 cells, or approximately 8 x 10 6 cells ~ approx. 9 x 10 6 For example, in one particular embodiment, the device can accommodate approximately 8.5 x 10 cells. 6 In some embodiments, the device can accommodate approximately 1 x 10 cells. 7 cells ~ approx. 1 x 108 In some embodiments, the device can accommodate approximately 1 x 10 cells. 8 cells ~ approx. 1 x 10 9 In some embodiments, the device can accommodate approximately 1 x 10 cells. 9 cells ~ approx. 1 x 10 10 In some embodiments, the device can accommodate approximately 1 x 10 cells. 10 cells ~ approx. 1 x 10 11 In some embodiments, the device can accommodate approximately 1 x 10 cells. 11 cells ~ approx. 1 x 10 12 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 x 10 12 In some embodiments, the device can accommodate approximately 1 x 10 cells after implantation in a subject. 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 x 10 12 Load cells.

[0174] The implantation device can be administered to a subject, for example, through a surgical incision at the site of implantation in the subject. The site of implantation can vary depending on the configuration of the device, the disorder being treated, and the desired biodistribution of biomolecules secreted by cells within the device. In some embodiments, the device is administered to a subject by implantation at a site selected from under the skin (subcutaneous implantation), on the omentum, or in or adjacent to the liver. In some embodiments, the cell chamber device is implanted on or under the skin; a mucosal surface, a body cavity, the peritoneal cavity; the central nervous system, e.g., the brain, cortex, ventricles, or spinal cord; an organ, e.g., the heart, liver, kidney, spleen, lung, pancreas, lymphatic system, vascular system, oral cavity, nasal cavity, teeth, gums, GI tract; bone; hip; adipose tissue; muscle tissue; circulatory blood; eye (e.g., intraocular); breast; vagina; uterus; a joint, e.g., the knee or hip joint, or the spine. In some embodiments, the cell chamber device is implanted in a specific part or tissue of the body, such as the blood, eye, brain, skin, lung, stomach, mouth, ear, leg, foot, hand, liver, heart, kidney, bone, reproductive organ, testicle, pancreas, spleen, large intestine, small intestine, spinal cord, or muscle. In some embodiments, the cell chamber device is administered to deliver biomolecules to a specific system of the body, such as the vascular system, nervous system (e.g., peripheral nervous system (PNS) or central nervous system (CNS)), skeletal system, respiratory system, endocrine system, lymphatic system, reproductive system, or gastrointestinal tract. In some embodiments, the cell chamber device is implanted under the subject's skin, on the subject's omentum, in the subject's subcutaneous fat, or in or adjacent to the subject's muscle tissue. In an exemplary embodiment, the device can be implanted subcutaneously.

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

[0176] The cell chamber device can be implanted in some cases to be easily retrieved from a subject, e.g., without injuring the subject or causing significant disruption of surrounding tissue. In one embodiment, the device can be retrieved with minimal or no surgical separation of the device from surrounding tissue, e.g., via a minimally invasive surgical approach, extraction, or excision.

[0177] The length of time the implant remains in the subject will vary depending on the disease being treated and the therapeutic dosage of the biomolecule secreted by the cells in the device. In some embodiments, the cell chamber device remains in the subject for at least 2 days, 3 days, 4 days, 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, 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, 11 years, 12 years, 13 years, 14 years, 15 years, or more. In some embodiments, the cell chamber device is maintained in the subject for about 1 to 2 days, 1 to 7 days, 2 to 4 days, 3 to 5 days, 4 days to 1 week, 1 to 2 weeks, 1 to 4 weeks, 2 to 4 weeks, 2 to 6 weeks, 3 to 5 weeks, 4 weeks to 1 month, 1 to 2 months, 1 to 4 months, 1 to 6 months, 2 to 4 months, 3 to 5 months, 4 to 6 months, 5 to 7 months, 6 to 8 months, 6 months to 9 months, 9 months to 10 months, 10 months to 12 months, 10 months to 14 months, 10 months to 16 months, 10 months to 18 ... The device may be maintained for an average period of 12 months, 7 months to 9 months, 8 months to 10 months, 9 months to 11 months, 10 months to 12 months, 11 months to 13 months, 12 months to 15 months, 15 months to 18 months, 18 months to 21 months, 21 months to 24 months, 2 years to 2.5 years, 2.5 years to 3 years, 3 years to 3.5 years, 3.5 years to 4 years, 4 years to 5 years, 5 years to 7 years, 7 years to 9 years, 9 years to 11 years, 11 years to 13 years, or 13 years to 15 years. In certain embodiments, the device is maintained 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. In certain embodiments, the device is implanted in the subject for at least 1 year. In certain embodiments, the device is implanted in the subject for at least 2 years. In certain embodiments, the device is implanted in the subject for at least 3 years. In certain embodiments, the device is implanted in the subject for at least 5 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 some embodiments, the device is permanently implanted in the subject.

[0178] In some embodiments, the cell chamber device is maintained in the subject such that the daily dose of the biomolecule (e.g., recombinant peptide or recombinant protein) secreted by the cells 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 more than 9 mg / day) for a period of time effective to achieve a therapeutic benefit in the subject (e.g., achieve a therapeutically effective plasma concentration of the biomolecule in the subject). In some embodiments, the cell chamber device is maintained in the subject such that the daily dose of the biomolecule (e.g., recombinant peptide or recombinant protein) secreted by the cells is between 0.5 mg / day and 1 mg / day, between 1 mg / day and 2 mg / day, between 1 mg / day and 5 mg / day, between 2 mg / day and 5 mg / day, between 4 mg / day and 7 mg / day, between 5 mg / day and 8 mg / day, between 5 mg / day and 10 mg / day, between 6 mg / day and 9 mg / day, between 7 mg / day and 10 mg / day, between 10 mg / day and 15 mg / day, between 8 mg / day and 11 mg / day, between 6 mg / day and 12 mg / day, or between 9 mg / day and 12 mg / day for a period of time effective to achieve a therapeutic benefit in the subject (e.g., to achieve a therapeutically effective plasma concentration of the biomolecule in the subject).

[0179] For example, in some embodiments, the cell chamber device is configured to increase the plasma concentration of a biomolecule (e.g., a recombinant peptide or recombinant protein) in a subject for a period of time after implantation, e.g., a period of time effective to achieve a therapeutic benefit in the subject, e.g., 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 5 months, at least 6 months, at least 12 months, 1 to 2 days, 1 to 7 days, 2 to 4 days, 3 to 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, ... and maintained in the subject to be 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) for 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 11 months, 10 months to 12 months, 11 months to 13 months, 12 months to 15 months, 15 months to 18 months, 18 months to 21 months, 21 months to 24 months, 2 years to 2.5 years, 2.5 years to 3 years, 3 years to 3.5 years, or 3.5 years to 4 years.

[0180] In some embodiments, the cell chamber device may be configured to increase the plasma concentration of the biomolecule (e.g., recombinant peptide or recombinant protein) in a subject over a period of time post-implantation, e.g., a period of time effective to achieve a therapeutic benefit in the subject, e.g., 1 to 2 days, 1 to 7 days, 2 to 4 days, 3 to 5 days, 4 days to 1 week, 1 to 2 weeks, 1 to 4 weeks, 2 to 4 weeks, 2 to 6 weeks, 3 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 12 months, 12 months to 14 months, 14 months to 16 months, 16 months to 18 months, 18 months to 20 months, 18 months to 22 months, 18 months to 24 months, 18 months to 26 months, 18 months to 28 months, 18 months to 29 months, 19 months to 20 months, 20 months to 30 months, 20 months to 31 months, 20 months to 32 months, 20 months to 33 months, 20 months to 34 months, 20 months to 35 months, 20 months to 36 months, 20 months to 37 months, 20 months to 38 months, 20 months to 39 months, 21 months to 34 months, 21 months to 35 months, 21 months to 36 months, 21 months to 38 months, 21 months to 39 months, 22 months to 39 months, and maintained in the subject at 4 μg / mL to 11 months, 10 months to 12 months, 11 months to 13 months, 12 months to 15 months, 15 months to 18 months, 18 months to 21 months, 21 months to 24 months, 2 years to 2.5 years, 2.5 years to 3 years, 3 years to 3.5 years, or 3.5 years to 4 years, 4 μg / mL to 8 μg / mL, 5 μg / mL to 9 μg / mL, 5 μg / mL to 12 μg / mL, 6 μg / mL to 10 μg / mL, 7 μg / mL to 11 μg / mL, 7 μg / mL to 15 μg / mL, 8 μg / mL to 12 μg / mL, 9 μg / mL to 13 μg / mL, 10 μg / mL to 15 μg / mL, 15 μg / mL to 20 μg / mL, or 20 μg / mL to 25 μg / mL.

[0181] In some embodiments, the cell chamber device may be configured to increase the plasma concentration of a biomolecule (e.g., a recombinant peptide or recombinant protein) in a subject after implantation, e.g., 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 5 months, at least 6 months, at least 12 months, 1 to 2 days, 1 to 7 days, 2 to 4 days, 3 to 5 days, 4 days to 1 week, 1 to 2 weeks, 1 week to 4 weeks, 2 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 12 months, ~11 months, 10 months to 12 months, 11 months to 13 months, 12 months to 15 months, 15 months to 18 months, 18 months to 21 months, 21 months to 24 months, 2 years to 2.5 years, 2.5 years to 3 years, 3 years to 3.5 years, or 3.5 years to 4 years, 1μg / mL to 5μg / mL, 1μg / mL to 10μg / mL, 5μg / mL to 10μg / mL, 5μg / mL to 15μg / mL, 10μg / mL to 15μg / mL, 1 0μg / mL~20μg / mL, 15μg / mL~20μg / mL, 15μg / mL~30μg / mL, 20μg / mL~25μg / mL, 20μg / mL~40μg / mL, 25μg / mL~30 µg / mL, 25 µg / mL to 50 µg / mL, 30 µg / mL to 35 µg / mL, 30 µg / mL to 60 µg / mL, or 35 µg / mL to 40 µg / mL.

[0182] In some embodiments, the cell chamber device is configured to increase the plasma concentration of a biomolecule (e.g., a recombinant peptide or recombinant protein) in a subject over a period of time following implantation, such as 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 5 months, at least 6 months, at least 12 months, 1-2 days, 1-7 days, 2-4 days, 3-5 days, 4-7 days, 7-10 days, 10-20 days, 20-30 days, 30-45 days, 45-60 days, 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 11 months, 10 months to 12 months, 11 months to 13 months, 12 months to 15 months, 15 months to 18 months, 18 months to 21 months, 21 months to 24 months, 2 years to 2.5 years, 2.5 years to 3 years, 3 years to 3.5 years, or 3.5 years to 4 years, and maintained in the subject to be at least 17 μg / mL.

[0183] In some embodiments, the cell chamber device is maintained within the subject for a period effective to deliver a dose of biomolecule of 0.1 mg / kg to about 10.0 mg / kg of the subject's body weight, e.g., about 2 mg / kg to about 7 mg / kg, about 3 mg / kg to about 6 mg / kg, or about 15 mg / kg to about 5 mg / kg. In certain embodiments, the dose of biomolecule delivered over the period of implantation is about 0.3 mg / kg, about 0.5 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, or about 10 mg / kg.

[0184] In some embodiments, the total dose of biomolecule delivered by the implant over a period of time, e.g., weekly, every two weeks, every four weeks, every six weeks, every eight weeks, or every ten weeks, may be about 22 mg, about 50 mg, about 72 mg, about 100 mg, about 125 mg, about 150 mg, about 165 mg, about 200 mg, about 300 mg, about 432 mg, about 450 mg, or about 600 mg. In some embodiments, the total dose of biomolecule delivered by the implant over a period of time, e.g., weekly, every two weeks, every four weeks, every six weeks, every eight weeks, or every ten weeks, may be, for example, at least 77 mg, at least 125 mg, or at least 356 mg. In one embodiment, the total dose of biomolecule delivered by the implant over a period of time, e.g., weekly, every two weeks, every four weeks, every six weeks, every eight weeks, or every ten weeks, is 165 mg. In another embodiment, the total dose every two weeks is 108 mg. In another embodiment, the total dose every two weeks is 216 mg. In another embodiment, the total dose every two weeks is 150 mg. In another embodiment, the total dose every two weeks is 200 mg. In another embodiment, the total dose every four weeks is 300 mg. In another embodiment, the total dose every eight weeks is 300 mg. For example, to deliver 300 mg of a biomolecule, e.g., an antibody, such as an anti-α4β7 antibody (e.g., vedolizumab), or an antigen-binding portion thereof, per eight-week period, the implant is sized and loaded with cells capable of delivering approximately 37.5 mg / week. In another example, to deliver 108 mg of a biomolecule, e.g., an antibody, such as an anti-α4β7 antibody (e.g., vedolizumab), or an antigen-binding portion thereof, per two-week period, the implant is sized and loaded with cells capable of delivering approximately 54 mg / week. The foregoing examples are illustrative only and are not intended to be limiting.

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

[0186] Where the biomolecule is an antibody, e.g., an anti-α4β7 antibody (e.g., vedolizumab), or an antigen-binding portion thereof, in some embodiments, the cell chamber device can be maintained in the subject such that the daily dose of the antibody, or antigen-binding portion thereof, secreted by the cells is about 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 more than 9 mg / day) for a period of time effective 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, or achieve a clinical benefit in the subject). In some embodiments, the cell chamber device is configured to maintain serum or plasma concentrations of the antibody, or antigen-binding portion thereof, secreted by the cells for a period of time effective to achieve a therapeutic benefit in the 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, at least 120 days, at least 150 days, at least 240 days, at least 365 days, or between 30 days, 90 days, 40 days, or 50 days) after implantation. and maintained in the subject such that the IL-16 concentration is 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 the subject for a period of at least 60 days, 50 days to 70 days, 60 days to 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 days to 240 days, 240 days to 365 days, or more than 365 days).

[0187] In some embodiments, the cell chamber device is maintained in the subject such that the daily dose of the antibody, or antigen-binding portion thereof, secreted by the cells is between 0.5 mg / day and 1 mg / day, between 1 mg / day and 2 mg / day, between 1 mg / day and 5 mg / day, between 2 mg / day and 5 mg / day, between 2 mg / day and 25 mg / day, between 4 mg / day and 7 mg / day, between 4 mg / day and 12 mg / day, between 5 mg / day and 8 mg / day, between 5 mg / day and 10 mg / day, between 6 mg / day and 9 mg / day, between 7 mg / day and 10 mg / day, between 10 mg / day and 15 mg / day, between 8 mg / day and 11 mg / day, between 6 mg / day and 12 mg / day, or between 9 mg / day and 12 mg / day in the subject for a period effective to achieve a therapeutic benefit in the subject.

[0188] In some embodiments, the cell chamber device is configured such that serum or plasma concentrations of the antibody, or antigen-binding portion thereof, secreted by the cells remain elevated for a period of time effective to achieve a therapeutic benefit in the 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, at least 120 days, at least 150 days, at least 240 days, at least 365 days, or between 30 and 90 days, 40 and 60 days, 50 and 70 days, 60 and 70 days, 60 and 90 days, 60 and 120 days, 65 and 75 days, and maintained in the subject at 2.5 μg / mL to 7.5 μg / mL, 4 μg / mL to 7 μg / mL, 5 μg / mL to 8 μg / mL, 5 μg / mL to 10 μg / mL, 5 μg / mL to 50 μg / mL, 8 μg / mL to 15 μg / mL, 10 μg / mL to 20 μg / mL, 16 μg / mL to 20 μg / mL, 18 μg / mL to 21 μg / mL, 21 μg / mL to 30 μg / mL, 25 μg / mL to 35 μg / mL, or 31 μg / mL to 45 μg / mL for a period of 70 to 90 days, 90 to 120 days, 120 to 240 days, 240 to 365 days, or more than 365 days.

[0189] In some embodiments, the cell chamber device is maintained within the subject for a period effective to deliver a dose of an antibody, e.g., an anti-α4β7 antibody (e.g., vedolizumab), or antigen-binding portion thereof, at about 0.1 mg / kg to about 10.0 mg / kg body weight, e.g., about 2 mg / kg to about 7 mg / kg, about 3 mg / kg to about 6 mg / kg, or about 15 mg / kg to about 5 mg / kg, about 5 mg / kg to about 25 mg / kg body weight, or about 10 mg / kg to about 20 mg / kg body weight. In certain embodiments, the total dose of the antibody (e.g., vedolizumab) or antigen-binding fragment thereof delivered over the duration of the implant is about 0.3 mg / kg, about 0.5 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, or about 10 mg / kg.

[0190] In some embodiments, the total dose of the antibody or antigen-binding fragment thereof delivered over the duration of the implant, e.g., about 2 weeks, about 4 weeks, about 6 weeks, about 8 weeks, about 10 weeks, about 12 weeks, about 14 weeks, about 16 weeks, about 20 weeks, about 24 weeks, about 28 weeks, about 32 weeks, about 36 weeks or more, can be about 22 mg, about 50 mg, about 72 mg, about 125 mg, about 165 mg, or about 432 mg, about 450 mg, about 300 mg, about 600 mg, about 650 mg, about 900 mg, about 1200 mg, about 1300 mg, about 1500 mg, about 1800 mg, about 2100 mg or more. In some embodiments, the total dose of the antibody or antigen-binding fragment thereof delivered over the duration of the implant can be at least 77 mg, at least 125 mg, or at least 356 mg. In one embodiment, the total dose of the antibody or antigen-binding fragment thereof delivered over the course of the implant is 165 mg. In another embodiment, the total dose of the antibody or antigen-binding fragment thereof is 108 mg. In another embodiment, the total dose of the antibody or antigen-binding fragment thereof is 216 mg. In another embodiment, the total dose of the antibody or antigen-binding fragment thereof is 300 mg. For example, an implant that stably delivers a total of three doses of a 300 mg biomolecule is typically administered intravenously every eight weeks, so that if it is implanted for 24 weeks, the device will deliver a total of 900 mg (about 5.36 mg per day). In another example, an implant that stably delivers a total of six doses of a 108 mg biomolecule is typically administered intravenously every two weeks, so that if it is implanted for 12 weeks, the device will deliver a total of 650 mg (about 7.7 mg per day).

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

[0192] In a further aspect, provided herein is a method of treating a subject having a disease by administering to the subject a device disclosed herein, wherein the cell chamber of the device contains cells that secrete a biomolecule in an amount and for a duration suitable to treat the disease or ameliorate one or more symptoms of the disease. Cells that secrete the desired biomolecule can be selected or engineered based on the disease being treated.

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

[0194] In some embodiments, the cell chamber device can be used to treat gastrointestinal disorders such as inflammatory bowel disease or short bowel syndrome. Examples of inflammatory bowel diseases (IBDs) that can be treated according to the methods herein include ulcerative colitis, Crohn's disease, primary sclerosing cholangitis, eosinophilic esophagitis, autoimmune hepatitis, ileitis, celiac disease, nontropical sprue, enteropathy associated with seronegative arthropathy, 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. In one embodiment, provided herein is a method of treating a subject with a gastrointestinal disorder, comprising administering to the subject a device disclosed herein. In certain such embodiments, a subject with a gastrointestinal disorder is administered a device containing cells that secrete a biomolecule that treats the gastrointestinal disorder.

[0195] In some embodiments, the cell chamber device is implanted in the peritoneum, mesentery, or peritoneal space of a subject with inflammatory bowel disease or short bowel syndrome. In some embodiments, the cell chamber device is implanted in the omentum, lesser omental bursa (i.e., omental bursa or omental pouch), greater omentum, in or above the lesser omentum, in or near the stomach, in or near the small intestine, in or near the large intestine, or small intestinal mesentery of a subject with inflammatory bowel disease or short bowel syndrome.

[0196] The cell chamber devices provided herein can be used to deliver to a subject any biomolecule secreted by cells encapsulated therein. For example, as described above, the cell chamber devices can be used to deliver protein or peptide therapeutics, as well as therapeutic antibodies, or antigen-binding portions thereof. By way of example, the following embodiments illustrate exemplary methods of treatment that can be achieved using the cell chamber devices described herein. These exemplary embodiments should not be construed as limiting the present invention in any way, as the cell chamber devices provided herein can be readily adapted to treat other diseases or disorders through the delivery of appropriate biomolecules, such as therapeutic proteins, peptides, antibodies, and the like.

[0197] In one embodiment, provided herein is a method of treating a subject with Crohn's disease or ulcerative colitis, comprising administering to the subject a device disclosed herein. In certain such embodiments, a subject with Crohn's disease or ulcerative colitis is administered a device comprising cells secreting an antibody, e.g., an anti-α4β7 antibody (e.g., vedolizumab or abrilumab), or an antigen-binding portion thereof. In some embodiments, the cell chamber device is configured to maintain serum or plasma concentrations of the antibody, or antigen-binding portion thereof, for a period effective to achieve a therapeutic benefit in the 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, 60-70 days, 60-90 days, 60-120 days) after implantation. and maintained in a subject with Crohn's disease or ulcerative colitis such that the serum erythrocyte serum concentration is 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 the subject for a period of at least 1 day, 65-75 days, 70-90 days, 90-120 days, 120-240 days, 240-365 days, or more than 365 days). In some embodiments, the cell chamber device is maintained within a subject with Crohn's disease or ulcerative colitis such that the device delivers the antibody, or antigen-binding portion thereof, at a rate of 10 mg / week to 50 mg / week, 20 mg / week to 60 mg / week, 30 mg / week to 75 mg / week, or 40 mg / week to 90 mg / week for a period effective to achieve a therapeutic benefit in the subject, e.g., at least 60 days (e.g., 60 to 70 days, 65 to 75 days, 70 to 90 days, 90 to 120 days, 120 to 240 days, 240 to 365 days, or more than 365 days) after implantation.In some embodiments, the cell chamber device is configured to provide a serum or plasma concentration of the antibody, or antigen-binding portion thereof, of between 2.5 μg / mL and 7.5 μg / mL, between 4 μg / mL and 7 μg / mL in the subject for a period of time effective to achieve a therapeutic benefit in the subject, e.g., at least 60 days (e.g., 60-70 days, 65-75 days, 70-90 days, 90-120 days, 120-240 days, 240-365 days, or more than 365 days) after implantation. In subjects with Crohn's disease or ulcerative colitis, the serum erythrocyte serum concentration is maintained at 100 μg / mL, 5 μg / mL to 8 μg / mL, 5 μg / mL to 10 μg / mL, 5 μg / mL to 50 μg / mL, 8 μg / mL to 15 μg / mL, 10 μg / mL to 20 μg / mL, 16 μg / mL to 20 μg / mL, 18 μg / mL to 21 μg / mL, 21 μg / mL to 30 μg / mL, 25 μg / mL to 35 μg / mL, or 31 μg / mL to 45 μg / mL. In some embodiments, subjects with Crohn's disease can have moderate to severe active Crohn's disease (e.g., a Crohn's Disease Activity Index (CDAI) score of 220 to 450). Treatment can achieve a clinical response or clinical remission in patients with moderate to severe active Crohn's disease. For example, treatment can result in mucosal healing. Treatment may also result in the reduction, elimination, or reduction and elimination of corticosteroid use by the patient (e.g., corticosteroid-free remission). In some embodiments, the subject with ulcerative colitis may have moderately to severely active ulcerative colitis (e.g., a Mayo score of 6-12 with an endoscopic subscore of 2-3). Treatment may result in the induction and maintenance of a clinical response, the induction and maintenance of clinical remission, or mucosal healing in patients with moderately to severely active ulcerative colitis. Treatment may also result in the reduction, elimination, or reduction and elimination of corticosteroid use by the patient (e.g., corticosteroid-free remission).

[0198] In another aspect, provided herein is a method of treating a subject with inflammatory bowel disease (IBD), comprising administering to the subject a cell chamber device disclosed herein, wherein cells in the cell chamber secrete an anti-α4β7 antibody (e.g., vedolizumab or abrilumab), or an antigen-binding portion thereof. Examples of inflammatory bowel diseases (IBD) that can be treated according to the methods herein include ulcerative colitis, Crohn's disease, ileitis, celiac disease, non-tropical sprue, enteropathy associated with seronegative arthropathy, microscopic or collagenous colitis, eosinophilic gastroenteritis, or pouchitis occurring after rectal resection, and ileoanal anastomosis. In some embodiments, the cell chamber device is configured to maintain serum or plasma concentrations of the antibody, or antigen-binding portion thereof, 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, 60-70 days, 60-9 ... and maintained in a subject with IBD such that the IBD level is 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 the subject for a period of time between 0 and 120 days, 65 and 75 days, 70 and 90 days, 90 and 120 days, 120 and 240 days, 240 and 365 days, or greater than 365 days).In some embodiments, the cell chamber device is configured to provide a serum or plasma concentration of the antibody, or antigen-binding portion thereof, of between 2.5 μg / mL and 7.5 μg / mL, 4 μg / mL, or more in the subject for a period of time effective to achieve a therapeutic benefit in the subject, e.g., at least 60 days (e.g., 60-70 days, 65-75 days, 70-90 days, 90-120 days, 120-240 days, 240-365 days, or more than 365 days) after implantation. The IBD concentration is maintained in subjects with IBD to be between 7 μg / mL, 5 μg / mL and 8 μg / mL, 5 μg / mL and 10 μg / mL, 5 μg / mL and 50 μg / mL, 8 μg / mL and 15 μg / mL, 10 μg / mL and 20 μg / mL, 16 μg / mL and 20 μg / mL, 18 μg / mL and 21 μg / mL, 21 μg / mL and 30 μg / mL, 25 μg / mL and 35 μg / mL, or 31 μg / mL and 45 μg / mL.

[0199] In some embodiments, the cell chamber device is implanted in the peritoneum, mesentery, or peritoneal space of a subject with Crohn's disease, ulcerative colitis, or other forms of IBD. In some embodiments, the cell chamber device is implanted in the omentum, lesser omental bursa (i.e., omental bursa or omental pouch), greater omentum, in or on the lesser omentum, in or near the stomach, in or near the small intestine, or in or near the large intestine of a subject with Crohn's disease, ulcerative colitis, or other forms of IBD.

[0200] In one embodiment, provided herein is a method of treating a subject with primary sclerosing cholangitis, comprising administering to the subject a device disclosed herein. In certain such embodiments, the subject with primary sclerosing cholangitis is administered a device comprising cells secreting an antibody, e.g., an anti-α4β7 antibody (e.g., vedolizumab or abrilumab), or an antigen-binding portion thereof. In some embodiments, the cell chamber device is administered a device containing cells secreting an antibody, e.g., an anti-α4β7 antibody (e.g., vedolizumab or abrilumab), or an antigen-binding portion thereof, for a period of time effective to achieve a therapeutic benefit in the 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, 60-70 days, 60-90 days, 60-90 days, 60-100 days, 60-110 days, 60-120 days, 60-130 days, 60-140 days, 60-150 days, 60-160 days, 60-170 days, 60-180 days, 60-190 days, 60-210 days, 60-220 days, 60-230 days, 60-240 days, 60-250 days, 60-260 days, 60-270 days, 60-280 days, 60-290 days, 60-300 days, 60-310 days, 60-320 days, 60-330 days, 60-340 days, 60-350 days, and maintained in a subject with primary sclerosing cholangitis such that the serum vasoconstriction is 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 the subject for a period of at least 120 days, 65 to 75 days, 70 to 90 days, 90 to 120 days, 120 to 240 days, 240 to 365 days, or more than 365 days).In some embodiments, the cell chamber device is configured to maintain serum or plasma concentrations of the antibody, or antigen-binding portion thereof, 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, 60-70 days, 60-90 days, 60-120 days, 65-75 days, 70-90 days, 90-120 days, 120-240 days, 240-36 days, 360-40 days, 360-50 days, 360-60 days, 360-70 days, 360-80 days, 360-90 days, 360-120 days, 360-140 days, 360-160 days, 360-240 days, 360-360 days, 360-40 days, 360-50 days, 360-60 days, 360-70 days, 360-80 days, 360-90 days, 360-140 days, 360-16 ... and maintained in a subject with primary sclerosing cholangitis such that the serum creatine phosphate level is between 2.5 μg / mL and 7.5 μg / mL, between 4 μg / mL and 7 μg / mL, between 5 μg / mL and 8 μg / mL, between 5 μg / mL and 10 μg / mL, between 5 μg / mL and 50 μg / mL, between 8 μg / mL and 15 μg / mL, between 10 μg / mL and 20 μg / mL, between 16 μg / mL and 20 μg / mL, between 18 μg / mL and 21 μg / mL, between 21 μg / mL and 30 μg / mL, between 25 μg / mL and 35 μg / mL, or between 31 μg / mL and 45 μg / mL in the subject for a period of 365 days or more (e.g., 5 days, or more than 365 days). In some embodiments, the cell chamber device is maintained in the subject with primary sclerosing cholangitis such that the device delivers the antibody, or antigen-binding portion thereof, at a rate of 10 mg / week to 50 mg / week, 20 mg / week to 60 mg / week, 30 mg / week to 75 mg / week, or 40 mg / week to 90 mg / week for a period effective to achieve a therapeutic benefit in the subject, e.g., at least 60 days (e.g., 60 to 70 days, 65 to 75 days, 70 to 90 days, 90 to 120 days, 120 to 240 days, 240 to 365 days, or more than 365 days) after implantation.

[0201] In some embodiments, the cell chamber device is implanted in the peritoneum, mesentery, or peritoneal space of a subject with primary sclerosing cholangitis. In some embodiments, the cell chamber device is implanted in the omentum, lesser omental bursa (i.e., omental bursa or omental sac), greater omentum, in or on the lesser omentum, in or near the liver, or in or near the sac of a subject with primary sclerosing cholangitis.

[0202] In one embodiment, provided herein is a method of treating a subject with lymphocytic esophagitis or eosinophilic esophagitis, comprising administering to the subject a device disclosed herein. In certain such embodiments, a subject with eosinophilic esophagitis is administered a device comprising cells secreting an antibody, e.g., an anti-α4β7 antibody (e.g., vedolizumab or abrilumab), or an antigen-binding portion thereof. In some embodiments, the cell chamber device is configured to maintain serum or plasma concentrations of the antibody, or antigen-binding portion thereof, for a period effective to achieve a therapeutic benefit in the 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, 60-70 days, 60-90 days, 60-120 days, and maintained in a subject with eosinophilic esophagitis (e.g., above the diaphragm) such that the eosinophilic esophagitis concentration is 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 the subject for a period of 65 to 75 days, 70 to 90 days, 90 to 120 days, 120 to 240 days, 240 to 365 days, or greater than 365 days).In some embodiments, the cell chamber device is configured to maintain serum or plasma concentrations of the antibody, or antigen-binding portion thereof, 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, 60-70 days, 60-90 days, 60-120 days, 65-75 days, 70-90 days, 90-120 days, 120-240 days, 240-30 days, 30-40 days, 40-50 days, 50-60 days, 60-70 days, 60-90 days, 60-120 days, 65-75 days, 70-90 days, 90-120 days, 120-240 days, 120-30 days, 120-40 days, 120-50 days, 120-60 days, 120-70 days, 120-80 days, 120-80 days, 120-90 days, 120-14 ... and the eosinophilic esophagitis concentration is maintained in a subject with eosinophilic esophagitis at 2.5 μg / mL to 7.5 μg / mL, 4 μg / mL to 7 μg / mL, 5 μg / mL to 8 μg / mL, 5 μg / mL to 10 μg / mL, 5 μg / mL to 50 μg / mL, 8 μg / mL to 15 μg / mL, 10 μg / mL to 20 μg / mL, 16 μg / mL to 20 μg / mL, 18 μg / mL to 21 μg / mL, 21 μg / mL to 30 μg / mL, 25 μg / mL to 35 μg / mL, or 31 μg / mL to 45 μg / mL in the subject for a period of 65 days or more (greater than 365 days). In some embodiments, the cell chamber device is maintained within a subject with lymphocytic esophagitis or eosinophilic esophagitis such that the device delivers the antibody, or antigen-binding portion thereof, at a rate of 10 mg / week to 50 mg / week, 20 mg / week to 60 mg / week, 30 mg / week to 75 mg / week, or 40 mg / week to 90 mg / week for a period effective to achieve a therapeutic benefit in the subject, e.g., at least 60 days (e.g., 60 to 70 days, 65 to 75 days, 70 to 90 days, 90 to 120 days, 120 to 240 days, 240 to 365 days, or more than 365 days) after implantation.

[0203] In some embodiments, the cell chamber device is implanted in the peritoneum, mesentery, or peritoneal space of a subject with eosinophilic esophagitis. In some embodiments, the cell chamber device is implanted in the omentum, lesser omental bursa (i.e., omental bursa or omental pouch), greater omentum, in or above the lesser omentum, in or near the esophagus, or above the diaphragm of a subject with lymphocytic esophagitis or eosinophilic esophagitis.

[0204] In one embodiment, provided herein is a method of treating a subject with autoimmune hepatitis, comprising administering to the subject a device disclosed herein. In certain such embodiments, the subject with autoimmune hepatitis is administered a device comprising cells secreting, for example, an antibody, an anti-α4β7 antibody (e.g., vedolizumab or abrilumab), or an antigen-binding portion thereof. In some embodiments, the cell chamber device is administered a device containing cells secreting an antibody, or an antigen-binding portion thereof, for a period of time effective to achieve a therapeutic benefit in the subject, 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, 30-90 days, 40-60 days, 50-70 days, 60-70 days, 60-90 days, 60 days). and maintained in a subject with autoimmune hepatitis such that the serum concentration is 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 the subject for a period of at least 120 days, 65 days to 75 days, 70 days to 90 days, 90 days to 120 days, 120 days to 240 days, 240 days to 365 days, or more than 365 days).In some embodiments, the cell chamber device is configured to maintain serum or plasma concentrations of the antibody, or antigen-binding portion thereof, 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, 60-70 days, 60-90 days, 60-120 days, 65-75 days, 70-90 days, 90-120 days, 120-240 days, 240-30 days, 30-40 days, 40-50 days, 50-60 days, 60-70 days, 60-90 days, 60-120 days, 65-75 days, 70-90 days, 90-120 days, 120-240 days, 120-30 days, 120-40 days, 120-50 days, 120-60 days, 120-70 days, 120-80 days, 120-80 days, 120-90 days, 120-14 ... and the serum creatine phosphate level is maintained in a subject with autoimmune hepatitis at 2.5 μg / mL to 7.5 μg / mL, 4 μg / mL to 7 μg / mL, 5 μg / mL to 8 μg / mL, 5 μg / mL to 10 μg / mL, 5 μg / mL to 50 μg / mL, 8 μg / mL to 15 μg / mL, 10 μg / mL to 20 μg / mL, 16 μg / mL to 20 μg / mL, 18 μg / mL to 21 μg / mL, 21 μg / mL to 30 μg / mL, 25 μg / mL to 35 μg / mL, or 31 μg / mL to 45 μg / mL in the subject for a period of 65 days, or more than 365 days. In some embodiments, the cell chamber device is maintained in the subject with autoimmune hepatitis such that the device delivers the antibody, or antigen-binding portion thereof, at a rate of 10 mg / week to 50 mg / week, 20 mg / week to 60 mg / week, 30 mg / week to 75 mg / week, or 40 mg / week to 90 mg / week for a period effective to achieve a therapeutic benefit in the subject, e.g., at least 60 days (e.g., 60 to 70 days, 65 to 75 days, 70 to 90 days, 90 to 120 days, 120 to 240 days, 240 to 365 days, or more than 365 days) after implantation.

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

[0206] Diseases or pathogens whose pathogenesis relies on the interaction of MAdCAM (e.g., MAdCAM-1) with α4β7 can also be treated with anti-α4β7 antibodies (e.g., vedolizumab or abrilumab), or antigen-binding portions thereof, delivered by the cell chamber device described herein. Examples of such diseases include immunodeficiency disorders such as those caused by human immunodeficiency virus (see, e.g., WO2008 / 140602).

[0207] Pancreatitis and insulin-dependent diabetes mellitus are other diseases that can be treated with anti-α4β7 antibodies (e.g., vedolizumab or abrilumab), or antigen-binding portions thereof, delivered using the cell chamber device disclosed herein. MAdCAM (e.g., MAdCAM-1) has been reported to be expressed by some blood vessels within the exocrine pancreas from NOD (non-obese diabetic) mice, as well as BALB / c and SJL mice. MAdCAM (e.g., MAdCAM-1) expression has been reported to be induced on the endothelium in inflamed pancreatic islets of NOD mice, and MAdCAM (e.g., MAdCAM-1) was the major addressin expressed by NOD islet endothelium during the early stages of insulitis (Hanninen, A., et al., J. Clin. Invest., 92:2509-2515 (1993)). Treatment of NOD mice with either anti-MAdCAM or anti-beta7 antibodies prevented the onset of diabetes (Yang et al., Diabetes, 46:1542-1547 (1997)). Furthermore, accumulation of α4β7-expressing lymphocytes was observed within pancreatic islets, and MAdCAM-1 was implicated in the binding of lymphoma cells to blood vessels from inflamed pancreatic islets via α4β7 in mantle cell lymphoma (Hanninen, A., et al., J. Clin. Invest., 92:2509-2515 (1993)) or to the gastrointestinal tract (Geissmann et al., Am. J. Pathol., 153:1701-1705 (1998)).

[0208] Other inflammatory diseases associated with mucosal tissue that may be treated with an anti-α4β7 antibody (e.g., vedolizumab or abrilumab), or antigen-binding portion thereof, delivered using the cell chamber device herein include cholecystitis, cholangitis (Adams and Eksteen Nature Reviews 6:244-251 (2006) Grant et al., Hepatology 33:1065-1072 (2001)), e.g., Behçet's disease, e.g., intestinal Behçet's disease, or pericholecititis (tissues surrounding the bile duct and liver), and graft-versus-host disease (e.g., in the gastrointestinal tract (e.g., after bone marrow implantation)) (Petrovic et al. Blood 103:1542-1547 (2004)). As seen in Crohn's disease, inflammation often extends beyond the mucosal surface; therefore, chronic inflammatory diseases such as sarcoidosis, chronic gastritis, e.g., autoimmune gastritis (Katakai et al., Int. Immunol., 14:167-175 (2002)), and other idiopathic conditions may be amenable to treatment.

[0209] Also provided herein are methods for inhibiting leukocyte infiltration of mucosal tissue using an anti-α4β7 antibody (e.g., vedolizumab or abrilumab), or an antigen-binding portion thereof, delivered using the cell chamber device herein. Further provided herein are methods for treating cancer (e.g., α4β7-positive tumors such as lymphoma). Other examples of inflammatory diseases associated with mucosal tissue that can be treated using an anti-α4β7 antibody (e.g., vedolizumab or abrilumab), or an antigen-binding portion thereof, delivered using the cell chamber device herein include mastitis (breast) and irritable bowel syndrome.

[0210] The cell chamber device can be loaded with several cells and implanted for a period of time sufficient to deliver an effective amount of an anti-α4β7 antibody, or antigen-binding portion thereof, that inhibits binding of α4β7 integrin to its ligand. With respect to therapy, an effective amount would be sufficient to achieve a desired therapeutic (including prophylactic) effect (e.g., an amount sufficient to reduce or prevent α4β7 integrin-mediated binding and / or signaling, thereby inhibiting leukocyte adhesion and infiltration and / or related cellular responses). An effective amount of anti-α4β7 antibody, e.g., an effective amount sufficient to maintain saturation, e.g., neutralization, of α4β7 integrin, can induce a clinical response or remission of the α4β7-associated conditions described herein, e.g., Crohn's disease, ulcerative colitis, inflammatory bowel disease, primary sclerosing cholangitis, eosinophilic esophagitis, autoimmune hepatitis, pancreatitis, cholecystitis, cholangitis, lymphoma, etc. The size of the cell chamber device and the number of anti-α4β7 antibody-producing cells within the device can be adjusted according to the methods of the present invention to produce an effective amount of anti-α4β7 antibody in a subject. For example, anti-α4β7 antibody can be delivered to a subject via the cell chamber device in the amounts and for the durations disclosed herein.

[0211] The dosage of anti-α4β7 antibody delivered by the cell chamber device can be optimized to induce clinical response and clinical remission in patients with inflammatory bowel disease. In some embodiments, the dosing regimen does not alter the CD4 to CD8 ratio in the cerebrospinal fluid of patients receiving treatment. The CD4:CD8 ratio can be measured in blood, lymph node aspirates, and cerebrospinal fluid (CSF). The CSF CD4+:CD8+ lymphocyte ratio in healthy individuals is typically about 1 or greater. (Svenningsson et al., J. Neuroimmunol. 1995;63:39-46; Svenningsson et al. Ann. Neurol. 1993;34:155-161). Immunomodulators can alter the CD4:CD8 ratio to less than 1.

[0212] In one embodiment, provided herein is a method of treating a subject with rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, psoriasis, hidradenitis suppurativa, uveitis, or juvenile idiopathic arthritis, comprising administering a device disclosed herein to the subject. In certain such embodiments, a subject with rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, psoriasis, hidradenitis suppurativa, uveitis, or juvenile idiopathic arthritis is administered a device comprising cells secreting an antibody, such as, for example, an anti-TNFα antibody (e.g., adalimumab, certolizumab, golimumab, or infliximab), or an antigen-binding portion thereof. In some embodiments, the cell chamber device is maintained in a subject with rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, psoriasis, psoriatic suppurative inflammation, utricle, or juvenile idiopathic arthritis such that the anti-TNFα antibody (e.g., adalimumab, certolizumab, golimumab, or infliximab), or an antigen-binding portion thereof, is secreted at a rate sufficient to provide a weekly, biweekly, monthly, or bimonthly dose of 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, at least 150 mg, at least 200 mg, at least 300 mg, or at least 400 mg) in the subject for a period effective to achieve a therapeutic benefit in the subject.In some embodiments, the cell chamber device is configured to deliver an anti-TNFα antibody (e.g., adalimumab, certolizumab, golimumab, or infliximab), or antigen-binding portion thereof, to a subject for a period of time effective to achieve a therapeutic benefit in the 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, 60-70 days, 60-90 days, 60-120 days, 65-75 days, 70-90 days, 90-120 days, 120-240 days, 240-365 days, or and maintained in a subject with rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, psoriasis, psoriatic suppurative arthritis, uveitis, or juvenile idiopathic arthritis such that the serum saturates are between 2.5 μg / mL and 7.5 μg / mL, 4 μg / mL and 7 μg / mL, 5 μg / mL and 8 μg / mL, 5 μg / mL and 10 μg / mL, 5 μg / mL and 50 μg / mL, 8 μg / mL and 15 μg / mL, 10 μg / mL and 20 μg / mL, 16 μg / mL and 20 μg / mL, 18 μg / mL and 21 μg / mL, 21 μg / mL and 30 μg / mL, 25 μg / mL and 35 μg / mL, or 31 μg / mL and 45 μg / mL in the subject for a period of 365 days or more.

[0213] In another aspect, provided herein is a method of treating a subject with inflammatory bowel disease (IBD), comprising administering to the subject a cell chamber device disclosed herein, wherein the cells in the cell chamber secrete an anti-TNFα antibody (e.g., adalimumab, certolizumab, golimumab, or infliximab). Examples of inflammatory bowel diseases (IBD) that can be treated according to the methods herein include ulcerative colitis, Crohn's disease, ileitis, celiac disease, non-tropical sprue, enteropathy associated with seronegative arthropathy, 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.

[0214] In another embodiment, provided herein is a method of treating a subject with ulcerative colitis or Crohn's disease, comprising administering to the subject a device disclosed herein. In certain such embodiments, the subject with ulcerative colitis or Crohn's disease is administered a device containing cells that secrete an antibody, such as an anti-integrin β7 antibody (e.g., etrolizumab). In some embodiments, the cell chamber device is maintained in the subject with ulcerative colitis or Crohn's disease such that the anti-integrin β7 antibody (e.g., etrolizumab), or an antigen-binding portion thereof, is secreted at a rate sufficient to provide a weekly, biweekly, monthly, or bimonthly dose of at least 50 mg (e.g., at least about 50 mg, at least about 25 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, at least about 225 mg, or at least about 250 mg) in the subject for a period effective to achieve a therapeutic benefit in the subject. In some embodiments, the cell chamber device is configured to maintain a plasma or serum concentration of the anti-integrin β7 antibody (e.g., etrolizumab), or antigen-binding portion thereof, 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, 60-70 days, 60-90 days, 60-120 days, 65-75 days, 70-90 days, 90-120 days, 120-240 days) after implantation. and / or maintained in a subject with ulcerative colitis or Crohn's disease such that the serum creatine phosphate level is between 2.5 μg / mL and 7.5 μg / mL, between 4 μg / mL and 7 μg / mL, between 5 μg / mL and 8 μg / mL, between 5 μg / mL and 10 μg / mL, between 5 μg / mL and 50 μg / mL, between 8 μg / mL and 15 μg / mL, between 10 μg / mL and 20 μg / mL, between 16 μg / mL and 20 μg / mL, between 18 μg / mL and 21 μg / mL, between 21 μg / mL and 30 μg / mL, between 25 μg / mL and 35 μg / mL, or between 31 μg / mL and 45 μg / mL in the subject for a period of 240 days to 365 days (240 days to 365 days, or more than 365 days).

[0215] In some embodiments, a subject with ulcerative colitis or Crohn's disease is administered a device containing cells that secrete an antibody, such as an anti-IL-12 / IL-23 antibody (eg, ustekinumab). In some embodiments, the cell chamber device is maintained in a subject with ulcerative colitis or Crohn's disease such that the anti-IL-12 / IL-23 antibody (e.g., ustekinumab), or antigen-binding portion thereof, is secreted at a rate sufficient to provide a weekly, biweekly, monthly, or bimonthly dose of 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 about 200 mg, at least about 225 mg, at least about 250 mg, at least about 300 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) in the subject for a period effective to achieve a therapeutic benefit in the subject. In some embodiments, the cell chamber device is configured to maintain plasma or serum concentrations of the anti-IL-12 / IL-23 antibody (e.g., ustekinumab), or antigen-binding portion thereof, 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, 60-70 days, 60-90 days, 60-120 days, 65-75 days, 70-90 days, 90-120 days, 120-24 days) after implantation. and maintained in a subject with ulcerative colitis or Crohn's disease such that the serum creatine phosphate level is between 2.5 μg / mL and 7.5 μg / mL, between 4 μg / mL and 7 μg / mL, between 5 μg / mL and 8 μg / mL, between 5 μg / mL and 10 μg / mL, between 5 μg / mL and 50 μg / mL, between 8 μg / mL and 15 μg / mL, between 10 μg / mL and 20 μg / mL, between 16 μg / mL and 20 μg / mL, between 18 μg / mL and 21 μg / mL, between 21 μg / mL and 30 μg / mL, between 25 μg / mL and 35 μg / mL, or between 31 μg / mL and 45 μg / mL in the subject for a period of 240 days, 240 days to 365 days, or more than 365 days.

[0216] In one embodiment, provided herein is a method of treating a subject with an autoimmune disease or cancer, the method comprising administering to the subject a device disclosed herein. In certain such embodiments, a subject with an autoimmune disease (e.g., rheumatoid arthritis) or cancer (e.g., non-Hodgkin's lymphoma, chronic lymphocytic leukemia) is administered a device comprising cells that secrete an antibody, such as an anti-CD20 antibody (e.g., rituximab). In some embodiments, the cell chamber device is configured to secrete an anti-CD20 antibody (e.g., rituximab) concentration, or the concentration of an antigen-binding portion thereof, of at least 200 mg / m 2 in the subject for a period of time effective to achieve a therapeutic benefit in the subject. 2 (e.g., at least about 200 mg / m 2 , at least about 225 mg / m 2 , at least about 250 mg / m 2 , 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 at least about 550 mg / m 2 In some embodiments, the cell chamber device is maintained in a subject with an autoimmune disease or cancer such that the anti-CD20 antibody (e.g., rituximab), or antigen-binding portion thereof, is administered at a concentration of at least 200-250 mg / m 2 in the subject for a period effective to achieve a therapeutic benefit in the subject. 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 / m 2 , 425-475 mg / m 2 , 450-500mg / m 2 , or 475-525 mg / m 2In some embodiments, the cell chamber device is maintained in a subject with an autoimmune disease or cancer such that the anti-CD20 antibody (e.g., rituximab), or an antigen-binding portion thereof, is secreted at a rate sufficient to provide a weekly, biweekly, monthly, or bimonthly dose of at least 150-200 mg, 175-225 mg, 200-250 mg, 225-275 mg, 250-325 mg, 300-350 mg, 325-375 mg, 350-400 mg, 375-425 mg, 400-450 mg, 425-475 mg, 450-500 mg, 475-525 mg, 500-550 mg, 525-575 mg, 550-600 mg, 575-600 mg, 580-600 mg, 590-700 mg, 600-750 mg, 610-750 mg, 620-750 mg, 630-750 mg, 640-750 mg, 650-700 mg, 660-700 mg, 670-750 mg, 680-700 mg, 690-800 mg, 710-720 mg, 730-740 mg, 750-750 mg, 760-770 mg, 780-790 mg, 790-800 mg, 800-810 mg, 810-820 mg, 820-830 mg, 830-840 mg, 840-850 mg, 850-860 mg, 860-870 mg, 870-880 mg, 880- and maintained in a subject with an autoimmune disease or cancer to be secreted at a rate sufficient to provide a weekly, biweekly, monthly, or bimonthly dose of 625 mg, 600-650 mg, 625-675 mg, 650-700 mg, 675-725 mg, 700-750 mg, 725-775 mg, 750-800 mg, 775-825 mg, 800-850 mg, 825-875 mg, 850-900 mg, 875-925 mg, 900-950 mg, 925-975 mg, 950-1000 mg, 900-1100 mg, or 1000-1200 mg.In some embodiments, the cell chamber device is configured to maintain plasma or serum concentrations of the anti-CD20 antibody (e.g., rituximab), or antigen-binding portion thereof, for a period of time effective to achieve a therapeutic benefit in the 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, 60-70 days, 60-90 days, 60-120 days, 65-75 days, 70-90 days, 90-120 days, 120-240 days, and maintained in a subject with an autoimmune disease or cancer such that the serum creatine mononitrate (SCN) is between 2.5 μg / mL and 7.5 μg / mL, between 4 μg / mL and 7 μg / mL, between 5 μg / mL and 8 μg / mL, between 5 μg / mL and 10 μg / mL, between 5 μg / mL and 50 μg / mL, between 8 μg / mL and 15 μg / mL, between 10 μg / mL and 20 μg / mL, between 16 μg / mL and 20 μg / mL, between 18 μg / mL and 21 μg / mL, between 21 μg / mL and 30 μg / mL, between 25 μg / mL and 35 μg / mL, or between 31 μg / mL and 45 μg / mL in the subject for a period of 240 days to 365 days, or more than 365 days.

[0217] In some embodiments, a subject with multiple sclerosis or Crohn's disease is administered a device containing cells that secrete an antibody, such as an anti-alpha-4 integrin antibody (e.g., natalizumab). In some embodiments, the cell chamber device is maintained within the subject with multiple sclerosis or Crohn's disease such that the anti-alpha-4 integrin antibody (e.g., natalizumab), or an antigen-binding portion thereof, is secreted at a rate sufficient to provide a weekly, biweekly, monthly, or bimonthly dose of 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, at least about 225 mg, at least about 250 mg, at least about 300 mg, or at least about 350 mg) in the subject for a period effective to achieve a therapeutic benefit in the subject. In some embodiments, the cell chamber device is configured to maintain a plasma or serum concentration of the anti-alpha-4 integrin antibody (e.g., natalizumab) or antigen-binding portion thereof 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, 60-70 days, 60-90 days, 60-120 days, 65-75 days, 70-90 days, 90-120 days, 120-240 days) after implantation. and maintained in a subject with multiple sclerosis or Crohn's disease such that the serum creatine mononitrate (SCN) is between 2.5 μg / mL and 7.5 μg / mL, between 4 μg / mL and 7 μg / mL, between 5 μg / mL and 8 μg / mL, between 5 μg / mL and 10 μg / mL, between 5 μg / mL and 50 μg / mL, between 8 μg / mL and 15 μg / mL, between 10 μg / mL and 20 μg / mL, between 16 μg / mL and 20 μg / mL, between 18 μg / mL and 21 μg / mL, between 21 μg / mL and 30 μg / mL, between 25 μg / mL and 35 μg / mL, or between 31 μg / mL and 45 μg / mL in the subject for a period of 240 days to 365 days (or more than 365 days).

[0218] In another embodiment, provided herein is a method of treating a subject with short bowel syndrome, comprising administering to the subject a device disclosed herein, hi certain such embodiments, a subject with short bowel syndrome is administered a device comprising cells that secrete a peptide therapeutic for gastrointestinal use.

[0219] In some embodiments, the cell chamber device can be used to treat a subject with cancer, such as melanoma, non-small cell lung cancer, small cell lung cancer, lung cancer, leukemia, liver cancer, retinoblastoma, astrocytoma, glioblastoma, gum 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 cancer, colon cancer, sarcoma, or bladder cancer. Thus, in one embodiment, provided herein is a method of treating a subject with cancer, comprising administering to the subject a cell chamber device disclosed herein. In certain such embodiments, a subject with cancer is administered a cell chamber device containing cells secreting a cancer-treating biomolecule (e.g., an antibody). For example, in some embodiments, the cells in the cell chamber device secrete an antibody, or an antigen-binding portion thereof, that specifically binds to CD20 (e.g., rituximab), VEGF (e.g., bevacizumab), HER2 / neu (e.g., trastuzumab), PD-L1 (e.g., atezolizumab, avelumab, durvalumab), PD-1 (e.g., pembrolizumab, nivolumab, cemiplimab), or EGFR (e.g., cetuximab, panitumumab).In some embodiments, the cell chamber device is configured to maintain plasma or serum concentrations of the antibody, or antigen-binding portion thereof, 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, 60-70 days, 60-90 days, 60-120 days, 65-75 days, 70-90 days, 90-120 days, 120-240 days, ... and maintaining the IL-14 concentration in the subject with cancer at 2.5 μg / mL to 7.5 μg / mL, 4 μg / mL to 7 μg / mL, 5 μg / mL to 8 μg / mL, 5 μg / mL to 10 μg / mL, 5 μg / mL to 50 μg / mL, 8 μg / mL to 15 μg / mL, 10 μg / mL to 20 μg / mL, 16 μg / mL to 20 μg / mL, 18 μg / mL to 21 μg / mL, 21 μg / mL to 30 μg / mL, 25 μg / mL to 35 μg / mL, or 31 μg / mL to 45 μg / mL for a period of 365 days or more (e.g., 365 days or more). In some embodiments, the implant can be placed subcutaneously near the site of the cancer in the subject, e.g., near an organ containing a tumor.

[0220] In some embodiments, the cell chamber device can be used to treat a subject with an autoimmune disease, such as graft-versus-host disease (GVHD), organ transplant rejection, autoimmune hepatitis, primary biliary cirrhosis, autoimmune 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 is a method of treating a subject with an autoimmune disease, comprising administering to the subject a cell chamber device disclosed herein. In certain such embodiments, a subject with an autoimmune disease is administered a cell chamber device containing cells that secrete a biomolecule that treats the autoimmune disease. For example, in some embodiments, the cells in the cell chamber device secrete an antibody, or an antigen-binding portion thereof, that specifically binds to α4β7 (e.g., vedolizumab, abrilumab), CD20 (e.g., rituximab), IL-12 / IL-23 (e.g., ustekinumab), integrin α4 (e.g., natalizumab), TNF-α (e.g., adalimumab, certolizumab, golimumab, infliximab), integrin β7 (e.g., etrolizumab), CD25 (e.g., basiliximab), IL-2Rα (e.g., daclizumab), or IgE (e.g., omalizumab).In some embodiments, the cell chamber device is configured to maintain plasma or serum concentrations 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 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, 60-70 days, 60-90 days, 60-120 days, 65-75 days, 70-90 days, 90-120 days, 120-240 days, 240-30 days, 30-40 days, 40-50 days, 50-6 ...50-70 days, 60-90 days, 60-120 days, 65-75 days, 70-90 days, 90-120 days, 120-240 days, 240-30 days, 30-40 days, 30-50 days, 40-60 days, 50-70 days, 60-70 days, 60-90 days, 60-120 days, 65-75 days, 70-90 days, 90-120 days, 120-240 days, 120-240 days, 120-30 days, 120-40 days, 120-50 days, 1 and maintaining the cell chamber device in a subject with an autoimmune disease at a concentration of 2.5 μg / mL to 7.5 μg / mL, 4 μg / mL to 7 μg / mL, 5 μg / mL to 8 μg / mL, 5 μg / mL to 10 μg / mL, 5 μg / mL to 50 μg / mL, 8 μg / mL to 15 μg / mL, 10 μg / mL to 20 μg / mL, 16 μg / mL to 20 μg / mL, 18 μg / mL to 21 μg / mL, 21 μg / mL to 30 μg / mL, 25 μg / mL to 35 μg / mL, or 31 μg / mL to 45 μg / mL in the subject for a period of 365 days or more. In some embodiments, the cell chamber device is implanted near lymphatic tissue or the mesentery.

[0221] In some embodiments, the cell chamber device can be used to treat a subject in need of enzyme replacement therapy (ERT). In such embodiments, the cell chamber can be seeded with cells expressing one or more enzymes that are deficient (e.g., absent, non-functional, partially functional, or expressed at suboptimal levels) in the subject. Thus, in some embodiments, provided herein is a method of treating a subject in need of enzyme replacement therapy, comprising administering to the subject a cell chamber device disclosed herein. In certain such embodiments, the device can contain cells that secrete the enzyme(s) that are deficient in the subject.

[0222] For example, in some embodiments, provided herein are methods of treating a subject with mucopolysaccharidosis type I (MPS I), comprising administering to the subject a device disclosed herein. In certain such embodiments, a subject with mucopolysaccharidosis type I (MPS I) is administered a device comprising cells that secrete an enzyme such as laronidase (e.g., SEQ ID NO: 50). In certain embodiments, the device comprises cells that secrete an enzyme comprising the amino acid sequence of SEQ ID NO: 50, or an enzyme having at least 90%, 92%, 94%, 95%, 96%, 98%, or 99% identity to SEQ ID NO: 50.

[0223] In other embodiments, provided herein are methods of treating a subject with mucopolysaccharidosis type II (MPS II), comprising administering to the subject a device disclosed herein. In certain such embodiments, a subject with mucopolysaccharidosis type I (MPS I) is administered a device comprising cells that secrete an enzyme such as idursulfase (e.g., SEQ ID NO: 51). In certain embodiments, the device comprises cells that secrete an enzyme comprising the amino acid sequence of SEQ ID NO: 51, or an enzyme having at least 90%, 92%, 94%, 95%, 96%, 98%, or 99% identity to SEQ ID NO: 51.

[0224] In other embodiments, provided herein are methods of treating a subject with metachromatic leukodystrophy (MLD), comprising administering to the subject a device disclosed herein. In certain such embodiments, a subject with mucopolysaccharidosis type I (MPS I) is administered a device comprising cells that secrete an enzyme such as arylsulfatase A (e.g., SEQ ID NO: 52). In certain embodiments, the device comprises cells that secrete an enzyme comprising the amino acid sequence of SEQ ID NO: 52, or an enzyme having at least 90%, 92%, 94%, 95%, 96%, 98%, or 99% identity to SEQ ID NO: 52.

[0225] In some embodiments, the cell chamber device can be used to treat a subject with a lysosomal storage disease, such as Pompe disease, adult-onset glycogen storage disease II (GSD II), Gaucher disease, Fabry disease, mucopolysaccharidosis type I, mucopolysaccharidosis type II, Niemann-Pick disease (including types A, B, and C), Morquio disease (including types A and B), Batten disease, Maroteaux-Lamy disease, metachromatic leukodystrophy, Tay-Sachs disease, sphingolipid disease, Hurler disease, or Hunter syndrome. Optionally, the lysosomal storage disease being treated is characterized by reduced or absent activity of a lysosomal enzyme in the patient's brain. Thus, in one embodiment, provided herein is a method of treating a subject with a lysosomal storage disease, comprising administering to the subject a cell chamber device disclosed herein. In certain such embodiments, a subject with a lysosomal storage disease is administered a cell chamber device containing cells secreting a biomolecule that treats the lysosomal storage disease. For example, in some embodiments, cells within the cell chamber device secrete enzymes that are deficient or absent in subjects with lysosomal storage diseases, including, but not limited to, hexosaminidase A, alpha-galactosidase A, glucocerebrosidase, arylsulfatase A, galactocerebrosidase, and sphingomyelinase. In some embodiments, cells within the cell chamber device secrete an enzyme therapeutic selected from agalsidase beta, agalsidase alfa, imiglucerase, taliglucerase alfa, velaglucerase alfa, alglucerase, sebelipase alfa, laronidase, idursulfase, elosulfase alfa, galsulfase, and alglucosidase alfa. The cell chamber devices and related methods disclosed herein are also useful for treating lysosomal storage disorders characterized by severe brain damage. In some embodiments, the cell chamber device delivers biomolecules across the blood-brain barrier. In other embodiments, the cell chamber device is implanted into the brain of a subject.

[0226] In some embodiments, the cell chamber device can be used to treat endocrine and / or metabolic disorders, such as diabetes, thyroid disorders, or osteoporosis. In one embodiment, provided herein is a method of treating a subject with an endocrine and / or metabolic disorder (e.g., diabetes), comprising administering to the subject a cell chamber device disclosed herein. In certain such embodiments, a subject with an endocrine and / or metabolic disorder (e.g., diabetes) is administered a cell chamber device comprising cells that secrete a biomolecule (e.g., insulin) that treats the endocrine and / or metabolic disorder. For example, in some embodiments, a cell chamber device for treating diabetes comprises cells that secrete insulin. In other embodiments, a cell chamber device for treating thyroid disorders comprises cells that secrete a thyroid hormone, e.g., levothyroxine. In other embodiments, a cell chamber device for treating osteoporosis comprises cells that secrete an antibody, or an antigen-binding portion thereof, that specifically binds to RANKL (e.g., denosumab).

[0227] In some embodiments, the cell chamber device can be used to treat blood disorders such as leukemia, lymphoma, myeloma, anemia, sickle cell anemia, or cachexia. In one embodiment, provided herein is a method of treating a subject with a blood disorder, comprising administering to the subject a cell chamber device disclosed herein. In certain such embodiments, the subject with a blood disorder is administered a cell chamber device containing cells that secrete a biomolecule that treats the blood disorder. For example, in some embodiments, the cells in the cell chamber device secrete an antibody, or an antigen-binding portion thereof, that specifically binds to CD20 (e.g., rituximab, obinutuzumab, ofatumumab), CD52 (e.g., alemtuzumab), CD19 (e.g., blinatumomab), CD22 (e.g., inotuzumab), CD38 (e.g., daratumumab), CD33 (e.g., gemtuzumab), or SLAMF7 (e.g., elotuzumab).

[0228] In some embodiments, the cell chamber device can be used to treat anemia, e.g., anemia associated with chronic kidney disease or end-stage renal disease. In certain such embodiments, a subject with anemia is administered a cell chamber device containing cells that secrete recombinant erythropoietin (e.g., epoetin alfa, dalpoietin alfa).

[0229] In some embodiments, the cell chamber device can be used to treat cardiovascular diseases such as congestive heart failure, hypertension, cardiomyopathy, myocarditis, atherosclerosis, chronic venous disease, or cardiac arrhythmia. In one embodiment, provided herein is a method of treating a subject with cardiovascular disease, comprising administering to the subject a cell chamber device disclosed herein. In certain such embodiments, a subject with cardiovascular disease is administered a cell chamber device containing cells that secrete a known biomolecule that treats the cardiovascular disease.

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

[0231] In some embodiments, sustained clinical remission, e.g., clinical remission sustained through at least two, at least three, or at least four visits with a caregiver within a six-month or one-year period after initiation of treatment, can be achieved following treatment with a cell chamber device herein. In some embodiments, sustained clinical response, e.g., clinical response sustained for at least six months, at least nine months, or at least one year after initiation of treatment, can be achieved following treatment with a cell chamber device herein.

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

[0233] Example 1: Fabrication of electrospun cell chambers The following method was used to construct a cell chamber device using electrospun polymer. This exemplary cell chamber device contains three nanofibrous electrospun polymer layers and one central polyester membrane. The following method can be adapted to fabricate devices containing additional electrospun polymer layers, fewer electrospun polymer layers, or different electrospun polymer layer(s). The method can also be adapted to fabricate devices lacking the central polyester membrane, if desired.

[0234] A first borosilicate vial was prepared containing polyethylene terephthalate (PET) and polybutylene terephthalate (PBT) chips and hexafluoroisoproposal (HFIP). A second borosilicate vial was prepared containing polyurethane (PU) chips (either polycarbonate, polyether, or polyester-based) and HFIP. The vials were placed on a rotator set at 45 RPM or higher until the chips were dissolved.

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

[0236] The film was removed from the mandrel and turned over so that the uncoated side was facing outward. A layer of nanofibrous electrospun PET-PBT was applied to the uncoated side of the film. A syringe loaded with PU solution was connected to the Espin unit, and a layer of nanofibrous electrospun PU was applied to one side of the film, on top of the nPET-PBT layer. In this way, a tri-layer scaffold containing a central film / membrane (nPET-PBT / / polyester membrane / / nPET-PBT / / PU) was created. The mandrel was immersed in ethanol and then sonicated to rinse. The scaffold was then rinsed with distilled water, dried, and removed from the mandrel.

[0237] The scaffold was cut into 50 mm x 40 mm sheets. The size of the scaffold could be increased or decreased depending on the desired size of the cell chamber. The scaffold was folded in half with the nPU layer facing inward to form the inner surface of the cell chamber. Three edges were ultrasonically welded using a Sonobond SeamMaster unit. The remaining edges were trimmed to a point by cutting the scaffold diagonally from the center point of the unsealed edge to a position below each closed edge. The resulting edges were ultrasonically welded closed so that the cell chamber had five closed sides. The center point was clipped to create a small opening within the chamber that could serve as a cell port, which could be resealed after loading. The scaffold could be treated with ethyldiamine to impart a positive charge to the polymer or sodium hydroxide to impart a negative charge to the polymer. To reduce inflammation due to implantation trauma, the cell scaffold device could also be optionally loaded with tacrolimus, as described in Example 8.

[0238] Example 2: Development of Vedolizumab / Luc-ARPE-19 cells In the following examples, ARPE-19 cells were selected as an exemplary cell type for loading into the cell chamber device. In this example, the ARPE-19 cells were engineered to secrete vedolizumab (e.g., for delivery to a subject when loaded into the cell chamber device). The cells were also engineered to express luciferase, allowing for cell monitoring in vitro and in vivo.

[0239] Two expression constructs were introduced into ARPE-19 cells. One expression construct encoded the vedolizumab heavy chain, and the second expression construct encoded luciferase and the vedolizumab light chain. Both the vedolizumab light and heavy chains were under the control of the human elongation factor-1 alpha (EF-1a) constitutive promoter. A clone that expressed vedolizumab (without luciferase) was also engineered. The cells were then grown and tested for the level of vedolizumab expression (see Table 1). [Table 1]

[0240] A vedolizumab-ARPE19 clone expressing approximately 40 picograms / cell / day was isolated. Additionally, ARPE19 cells expressing both vedolizumab and luciferase were isolated for use in the Examples described further herein (vedolizumab / Luc-ARPE19 clone).

[0241] Vedolizumab secretion per cell per day and per cm of scaffold 2 A simulated model was prepared to estimate the plasma concentration levels of vedolizumab that could be achieved over time based on the estimated cell numbers per 625,000 cells / cm on the scaffold. 2 It was estimated that the cells would grow at 625,000 cells / cm 2 However, if 45 picograms of vedolizumab is secreted per day, this translates to approximately 28 μg of vedolizumab / cm per day. 2 would be secreted. Thus, an 8 x 10 cm device would secrete approximately 4.5 mg of vedolizumab / day. As shown in the simulation in Figure 2, such a device would be predicted to reach therapeutic plasma concentrations of vedolizumab (17 μg / mL) within 60 days of implantation.

[0242] Example 3: In vitro cell seeding assay with vedolizumab / Luc-ARPE-19 cells Vedolizumab and luciferase-expressing ARPE-19 cells (vedolizumab / Luc-ARPE19 cells) were seeded onto 16 mm discs consisting of an outer layer of polyethylene terephthalate-polybutylene terephthalate (nPET-PBT) and an inner layer of polyurethane (PU).

[0243] To evaluate whether cells could grow on the inner PU layer of the scaffold, dolizumab / Luc-ARPE19 cells were stained with CellTracker Orange (5 μM) and 250,000 pre-stained cells were seeded on the PU side of nPET-PBT / PU (16 mm discs). The discs were then observed by fluorescent 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. Furthermore, no cell migration from the PU side to the PBT-PET side was observed.

[0244] The number of cells loaded onto nPET-PBT / PU disks was compared with the number of cells that could be loaded onto tissue culture (TC) plates. Images of cells on tissue culture plates and nPET-PBT / PU disks are shown in Figure 3B. The top row shows CellTracker Orange fluorescence in the cytoplasm of cells in each condition, and the bottom row shows nuclei stained with Hoechst 33342. The number of cells on nPET-PBT / PU disks compared to tissue culture plates was determined by quantifying the amount of DNA isolated from cells in each condition. As shown in Figure 3C, the amount of DNA isolated from cells grown on nPET-PBT / PU was five times that isolated from cells on TC plates, indicating that at least five times more cells could be cultured on nPET-PBT / PU than on tissue culture plates. Furthermore, a comparison of the amount of vedolizumab secreted in each condition indicated that cells on nPET-PBT / PU membranes secreted five times more vedolizumab than cells on tissue culture plates (Figure 3D).

[0245] Vedolizumab / Luc-ARPE19 cell seeding and secretion of vedolizumab by the cells were then evaluated on nPET-PBT with charged surface modifications. Discs containing nPET-PBT / PU were treated with liquid ethylenediamine (nPET-PBT(EDA)) to generate a positively charged surface, or with liquid sodium hydroxide to generate a negatively charged surface (nPET-PBT(HYD)). ARPE19 clones expressing vedolizumab / luciferase were seeded onto discs containing nPET-PBT / PU, nPET-PBT, nPET-PBT(EDA), or nPET-PBT(HYD) at a density of 1 million cells per 16 mm disc. Cells were maintained in DMEM / F-12 medium supplemented with 10% FBS in a 37°C incubator with 5% CO2, and the medium was replaced with fresh medium every 3–4 days. Three weeks after seeding, vedolizumab secretion (Figure 4A) and luminescence intensity (Figure 4B) were assessed by vedolizumab ELISA and IVIS imaging, respectively. As shown in Figures 4A and 4B, vedolizumab / Luc-ARPE-19 cells were seeded on the surface-modified materials, and the cells were able to maintain vedolizumab secretion.

[0246] We then evaluated the cytokine secretion profile of vedolizumab / Luc-ARPE19 cells grown on charge-modified or unmodified nPET-PBT. Vedolizumab / Luc-ARPE19 cells were seeded on discs containing nPET-PBT / PU, nPET-PBT, nPET-PBT(EDA), or nPET-PBT(HYD). Three weeks after seeding, cytokine secretion by vedolizumab / Luc-ARPE19 cells was evaluated. Data were normalized by secretion data from cells seeded on nPET-PBT / PU membranes. As shown in Table 2, there was only a 0.5- to 1.2-fold change in cytokine secretion from ARPE-19 cells, indicating that the charge surface modification did not affect cell function. [Table 2]

[0247] Example 4: In vitro and in vivo cell distribution of vedolizumab / Luc-ARPE-19 cells in nanofibrous polymer cell chamber devices In this study, two cell chamber devices were tested. The first device contained a scaffold comprising an outer layer of nPET-PBT and an inner layer of PU surrounding the cell chamber. This device is shown in Figure 1C. The other device contained the same nPET-PBT / PU scaffold, plus a second scaffold positioned within the cell chamber, comprising pore-modified nanofibrous polybutylene (nPBT). This device is shown in Figure 1D. To evaluate cell distribution in each device in vitro, the cell chambers were loaded with vedolizumab / Luc-ARPE19 cells. Cell distribution was assessed by luminescence imaging 4 days after cell seeding. As shown in Figure 5A, cells were distributed across both devices, i.e., the nPET-PBT / PU cell chamber with and without the inner nPBT scaffold.

[0248] To evaluate cell viability and distribution on nPET-PBT / PU chambers with or without an inner nPBT scaffold after in vivo implantation, both chambers were implanted (subcutaneously, near the shoulder) into BALB / c nude mice. Mice were evaluated by luminescence imaging and blood sampling twice weekly over a 62-day period. Luminescence imaging was performed by intraperitoneally injecting D-luciferin / PBS solution (15 mg / mL, 100 μL / 10 g) and subsequently imaging the mice 10 minutes after injection. As shown in Figures 5B and 5C, which show the luminescence intensity from the implanted chambers as a function of days after implantation, cells survived in the cell chamber devices with or without the inner nPBT scaffold for 65 or 90 days, respectively. Furthermore, as shown in Figure 5D, vedolizumab was detected by Western blot (primary antibody: goat anti-human antibody, Fc region-specific; secondary antibody: rabbit anti-goat antibody, HRP conjugate) in mice implanted with either chamber device 90 days after implantation.

[0249] To assess whether cells had leaked from the chamber, D-luciferin was intraperitoneally injected into mice implanted with cell chambers, and IVIS imaging was performed 8 minutes after injection. The mice were then euthanized, the chambers were removed, and IVIS imaging was performed again to determine whether cells were located on the back of the mouse. As shown in Figure 5E, there was no detectable luminescence signal on the back of the mouse after chamber removal, indicating that vedolizumab / Luc-ARPE19 cells had not leaked from the chamber. The explanted chambers removed 42 days after implantation were evaluated. As shown in Table 3, the explanted chambers continued to secrete vedolizumab. Additionally, as shown in Figures 5F and 5G, no significant fibrotic response was visually observed. [Table 3]

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

[0251] Example 5: In vitro cell attachment assay with vedolizumab / Luc-ARPE-19 cells To evaluate cell attachment on the surface-modified materials, ARPE-19 cells engineered to express luciferase (Luc-ARPE-19 cells) were evaluated for cell attachment to disks composed of nPET-PBT, nPET-PBT(EDA), or nPET-PBT(HYD). Cell attachment to normal tissue culture (TC) plates was evaluated as a control. 400,000 cells were seeded per disk of material, and the percentage of attached cells was assessed at multiple time points after seeding.

[0252] As shown in Figure 6, ARPE-19 cells rapidly attached to all three electrospun surfaces in a time-dependent manner. The charged materials exhibited comparable attachment to tissue culture plastic, with all materials loading 50-60% of the cells within 60 minutes. However, the charged materials (nPET-PBT(EDA) or nPET-PBT(HYD)) had greater cell loading (90% attached cells) than unmodified nPET-PBT after 180 minutes of incubation.

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

[0254] Example 6: In vivo implant assay with vedolizumab / Luc-ARPE-19 cells To test cell retention on electrospun scaffolds in vivo, 16 mm membrane discs composed of nPET-PBT, nPET-PBT(EDA), or nPET-PBT(HYD) were seeded with cells stably expressing vedolizumab and luciferase (vedolizumab / Luc-ARPE19 cells; 400,000 cells per 16 mm disc) and implanted subcutaneously into female nude mice. The discs used in this experiment did not contain an enclosed chamber. Administration of 100 μl of vedolizumab / Luc-ARPE19 cells via subcutaneous injection was evaluated as a control. Cells were loaded into each disc on the day of implantation. Scaffolds were administered to mice by subcutaneous implantation 24 or 4 hours after loading (n = 4 mice). Luminescence was monitored by live-cell imaging every 3–4 days over a 70-day period to determine the extent of cell retention. As shown in Figures 7A-7E, ARPE-19 cells on electrospun materials had a longer retention time at the implantation site than subcutaneously injected cells.

[0255] Example 7: In vitro cell seeding assay with vedolizumab / Luc-ARPE-19 cells on tacrolimus-treated nPET-PBT To reduce inflammation due to implantation trauma, cell scaffold devices can be loaded with tacrolimus (FK506). To evaluate cell seeding and biomolecule secretion on tacrolimus-treated materials, electrospun nPET-PBT was loaded with tacrolimus by dissolving 0%, 2%, or 4% tacrolimus in the polymer solution before electrospanning.

[0256] The incorporation of tacrolimus into nPET-PBT was evaluated by solvent extraction of tacrolimus from the electrospun material. The solvent-extracted solution was then analyzed by high-performance liquid chromatography (HPLC). As shown in Figure 8A (tacrolimus alone) and Figure 8B (tacrolimus after extraction), HPLC analysis indicated no drug degradation after loading of tacrolimus onto electrospun nPET-PBT.

[0257] To evaluate the biological activity of tacrolimus-loaded nPET-PBT, an in vitro T cell activation assay was performed in which T cells were incubated in medium exposed to tacrolimus-loaded nPET-PBT. Electrospun nPET-PBT was immersed in 400 μL of RPMI 1640 supplemented with 10% fetal bovine serum (FBS) and incubated for 24 hours (initial sample). The nPET-PBT was then transferred to pre-warmed fresh medium and incubated for 24 hours (second sample). The nPET-PBT was then transferred again to pre-warmed fresh medium and incubated for 24 hours (third sample). Each medium sample was collected for drug concentration evaluation by ELISA / HPLC. Human PBMCs were incubated with the collected medium sample for 30 minutes. Subsequently, the human PBMCs were activated with CD3 / CD28-activated Dynabeads for 48 hours. The supernatant from the T cell culture medium was then isolated and tested for IL-1β production. As shown in Figure 8C, medium exposed to tacrolimus-loaded nPET-PBT inhibited T cell activation as measured by IL-1β levels, indicating that tacrolimus remained effective after loading into the electrospun material and that tacrolimus was gradually released over a 3-day period (Figure 8C).

[0258] Next, an in vitro assay was performed to evaluate whether tacrolimus-loaded electrospun nPET-PBT affected cell growth and vedolizumab secretion by vedolizumab / Luc-ARPE19 cells. Vedolizumab / Luc-ARPE19 cells (200,000 cells / state) were seeded onto nPET-PBT with or without tacrolimus. Cells were grown on the tacrolimus-loaded nPET-PBT disks for 3 days, after which 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 in Figures 8E and 8F, vedolizumab / Luc-ARPE-19 cells seeded onto tacrolimus-loaded nPET-PBT were able to maintain vedolizumab secretion. These results indicate that tacrolimus had no off-target effects on vedolizumab / Luc-ARPE19 cells.

[0259] We then evaluated the cytokine secretion profile of vedolizumab / Luc-ARPE19 cells grown on nPET-PBT membranes with or without tacrolimus. Cells were grown on tacrolimus-treated nPET-PBT membranes as described above for 3 days. Cytokine secretion was assessed for cells seeded on each substrate 3 weeks after seeding. Data were normalized by secretion data from cells seeded on nPET-PBT membranes without tacrolimus treatment. As shown in Table 4, there was only a 0.5- to 2-fold change in cytokine secretion from ARPE-19 cells, indicating that tacrolimus did not affect cell function. [Table 4]

[0260] Example 8: In vitro cell seeding assay with adalimumab / ARPE-19 cells and ustekinumab / ARPE-19 cells ARPE-19 cells were generated to express and secrete adalimumab (adalimumab / ARPE19 cells) or ustekinumab (ustekinumab / ARPE19 cells) in the same manner as described above for the generation of vedolizumab-secreting ARPE-19 cells.

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

[0262] The secretion of biomolecules (i.e., adalimumab or ustekinumab) by cells loaded onto nPET-PBT / PU membranes 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 nPET-PBT / PU membranes secreted more adalimumab (Figure 9A) or ustekinumab (Figure 9B) than cells on tissue culture plates.

[0263] Example 9: Functional activity of antibodies secreted using the cell chamber device The functional activity of antibodies secreted by ARPE-19 cells was evaluated by in vitro assay. ARPE-19 cells secreting adalimumab, ustekinumab, or vedolizumab (adalimumab-ARPE-19, ustekinumab-ARPE-19, and vedolizumab-ARPE-19) were seeded in cell culture flasks at a density of 50,000 cells / cm^2 in SFM4 MegaVir medium and cultured for 15 days. The amount of each antibody secreted by the cells was determined based on adalimumab ELISA, ustekinumab ELISA, and vedolizumab ELISA assays (see Tables 1 and 5). Cell culture medium containing the antibodies secreted by the cells was assayed for antibody functional activity as outlined below.

[0264] Adalimumab is an anti-TNFα antibody. To assay adalimumab activity in the cell culture medium of ARPE-19 cells, L-929 cells were seeded in a 96-well cell culture plate at a density of 5,000 cells / well in DMEM supplemented with 10% FBS. After 20 hours of incubation, the cells were sensitized with actinomycin D (2 μg / mL) for 2 hours. The cells were then incubated with culture medium containing the antibody secreted by ARPE-19 cells in the presence of human TNFα (1 ng / mL). After a further 20 hours of incubation, the cells were assessed for cell viability by CCK-8 assay and tested for neutralization of human TNFα activity. As shown in Figure 10A, these results indicate that adalimumab secreted by ARPE-19 cells retained TNFα neutralizing activity.

[0265] Vedolizumab is a humanized α4β7 integrin antibody. To assay for vedolizumab activity in the cell culture medium of three vedolizumab-expressing ARPE-19 clones and one vedolizumab / luciferase-expressing ARPE-19 clone, an α4β7 integrin binding assay was performed using HuT-78 cells, which specifically express α4β7 integrin, using flow cytometry. Cells were cultured in SFM4 MegaVir medium for 2 weeks, and these medium samples were used in the binding assay after determining vedolizumab concentration by vedolizumab ELISA. The binding activity of the conditioned medium was assessed by a competition assay using fluorescently labeled purified vedolizumab (R-PE-labeled MLN002) secreted from CHO cells. As sh...

Claims

1. A device comprising a multi-layered scaffold surrounding a cell chamber, the multi-layered 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 wherein the outer layer and / or the inner layer comprises one or more charged surface modifications.

2. the outer layer comprises electrospun nanofibrous polyethylene terephthalate and polybutylene terephthalate; The device of claim 1 .

3. the inner layer comprises electrospun polyurethane, or electrospun polyethylene terephthalate and polybutylene terephthalate; The device of claim 2 .

4. the outer layer and / or the inner layer have a net positive charge, or the outer layer and / or the inner layer have a net negative charge; 4. A device according to claim 2 or 3.

5. The device of claim 1 or 4, wherein the scaffold is treated with ethylenediamine and / or sodium hydroxide.

6. the outer layer and / or the inner layer comprises an anti-inflammatory agent; A device according to any one of claims 1 to 5.

7. the anti-inflammatory agent is selected from the group consisting of a calcineurin inhibitor, tacrolimus, pyridone, pirfenidone, a phosphodiesterase inhibitor, and roflumilast; The device of claim 6.

8. The device according to any one of claims 1 to 7, wherein the outer layer and / or the inner layer comprises pores.

9. The pores are sized to prevent the passage of cells, the pores having a diameter of 1 μm or less, or the pores having a diameter of 0.5 μm or less. The device of claim 8.

10. the multi-layered scaffold further comprising a porous membrane positioned between the inner layer and the outer layer. A device according to any one of claims 1 to 9.

11. the porous membrane comprises nanofibrous polyethylene terephthalate or non-nanofiberous polyethylene terephthalate; The device of claim 10.

12. The porous membrane contains pores of a size that allows the passage of a biomolecule, and the biomolecule is 250 kDa or less.

12. A device according to claim 10 or 11.

13. The device of claim 12, wherein the membrane pores have a diameter of 0.2 to 0.6 μm.

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

15. The device of any one of claims 1 to 14, further comprising a loading port that allows cells to be loaded into the cell chamber.

16. the device comprises a total thickness of 250 μm or less; A device according to any one of claims 1 to 15.

17. The cell chamber contains 1×10 ７ cells ~ 1 x 10 ９ It is sized to accommodate cells, A device according to any one of claims 1 to 16.

18. 4. The device of claim 2 or 3, wherein the multilayer scaffold comprises an outer layer comprising nanofibrous polyethylene terephthalate and polybutylene terephthalate and an inner layer comprising nanofibrous polyurethane, the outer layer and the inner layer comprising nanopores having a diameter of 1 μm or less.

19. the cell chamber contains cells; A device according to any one of claims 1 to 18.

20. 20. The device of claim 19, wherein the cells adhere to the inner layer of the scaffold.

21. 21. The device of claim 19 or 20, wherein the cells comprise retinal pigment epithelial cells.

22. the cells secrete the recombinant peptide or protein; A device according to any one of claims 19 to 21.

23. 23. The device of claim 22, wherein the cells secrete a protein selected from the group consisting of an antibody, or an antigen-binding portion thereof, a growth factor, a hormone, an enzyme, a cytokine, a peptide therapeutic, or a combination thereof.

24. the cells secrete a peptide therapeutic; 24. A device according to claim 22 or 23.

25. 25. The device of claim 24, wherein the cells secrete an antibody, or an antigen-binding portion thereof.

26. 26. The device of claim 25, wherein the antibody, or antigen-binding portion thereof, specifically binds to an antigen selected from the group consisting of α4β7, integrin β7, TNFα, IL-12, IL-23, or CD20.

27. 27. The device of claim 26, wherein the antibody, or antigen-binding portion thereof, that specifically binds to α4β7 is vedolizumab.

28. The cell chamber contains cells having a three-dimensional structure.

21. A device according to claim 19 or 20.

29. 29. The device of claim 28, wherein the cells having a three-dimensional structure comprise a tissue explant, or comprise an organoid or spheroid.

30. 30. The device of claim 29, wherein the organoids are organized around sinusoids or ducts.

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