Cell encapsulating device and method using the same

Abstract

To provide a method and a device for improving transplantation of a cell to a target which include improving viability and functionality of a transplantation cell.SOLUTION: A device for encapsulating a biological cell is suitable for being transplanted to a target. The device can include a plurality of polymer layers. The device includes a first polymer layer and a second polymer layer. In some cases, the first polymer layer can be a nanoporous polymer layer. In some cases, the second polymer layer can be a macroporous polymer layer. The first and second polymer layers can regulate a lumen for encapsulating population of cells. The device is usable for transplanting a cell producing a therapeutic agent (for example, for treatment of disease) to a target.SELECTED DRAWING: None

Description

[Technical field]

[0001] cross reference This application claims the benefit of U.S. Provisional Application No. 62 / 816,675, filed March 11, 2019, which is incorporated herein by reference in its entirety. [Background technology]

[0002] 2. Background of the Invention Transplantation of cells into a subject to generate therapeutic molecules can be an attractive alternative to the use of exogenous drugs. Encapsulation of cell transplants can, for example, prevent proliferation of transplanted cells in the host and increase the viability of transplanted cells. The present disclosure provides methods and devices for improving the transplantation of cells into a subject, including improving the viability and functionality of transplanted cells. Summary of the Invention [Means for solving the problem]

[0003] In one embodiment, a device is provided that includes: a) a first polymer layer comprising a plurality of pores having an average connection diameter of less than about 500 nm; and b) a second polymer layer comprising a plurality of pores having an average connection diameter of more than about 100 μm, wherein the first polymer layer and the second polymer layer define a lumen for encapsulating a plurality of cells. In some cases, the plurality of pores present in the first polymer layer have an average connection diameter of about 10 nm to about 200 nm. In some cases, the plurality of pores present in the first polymer layer have an average connection diameter of about 180 nm to about 220 nm. In some cases, the plurality of pores present in the second polymer layer have an average connection diameter of about 100 μm to about 500 μm. In some cases, the plurality of pores present in the second polymer layer have an average connection diameter of about 200 μm to about 400 μm. In some cases, the plurality of pores present in the first polymer layer have an average pore diameter of about 1 μm to about 5 μm. In some cases, the plurality of pores present in the second polymer layer has an average pore diameter of about 500 μm to about 3 mm. In some cases, the plurality of pores present in the second polymer layer has an average pore diameter of about 100 μm to about 1 mm. In some cases, the first polymer layer has a thickness of about 10 μm to about 200 μm. In some cases, the first polymer layer has a thickness of about 20 μm to about 50 μm. In some cases, the first polymer layer has a thickness of about 18 μm to about 22 μm. In some cases, the second polymer layer has a thickness of about 1 mm to about 5 mm. In some cases, the second polymer layer has a thickness of about 400 μm to about 600 μm. In some cases, the device further comprises a third polymer layer having an average connection diameter of less than about 500 nm. In some cases, the device further comprises a port for introducing a plurality of cells into the lumen of the device.In some cases, the first polymer layer, the second polymer, the third polymer layer, or any combination thereof comprises a biocompatible polymer selected from the group consisting of polycaprolactone (PCL), polyvinylidene fluoride (PVDF), polytetrafluoroethylene (PTFE), polyethylene (PE), methacrylate polymers, polyethyleneimine, polyethyleneimine-dextran sulfate, poly(vinylsiloxane) ecopolymer polyethyleneimine, phosphorylcholine, poly(ethyl methacrylate), polyurethane, poly(ethylene glycol) (PEG), poly(lactic-glycolic acid) (PLGA), hydroxyapatite, poly(lactic acid), polyhydroxyvaleric acid and its copolymers, polyhydroxybutyric acid and its copolymers, polydiaxanone, polyanhydrides, polycyanocrylate, poly(amino acids), poly(orthoesters), polyesters, collagen, gelatin, cellulose polymers, chitosan, alginate, laminin, and any combination thereof. In some cases, the biocompatible polymer is polycaprolactone. In some cases, the plurality of cells is selected from the group consisting of thyroid cells, parathyroid cells, bone marrow cells, mesenchymal stem cells, stromal cells, pluripotent stem cells, induced pluripotent stem cells, embryonic stem cells, vascular cells, cells derived from adipose tissue, cells derived from bone marrow, intestinal cells or cells derived therefrom, pancreatic islets or islet cells, Sertoli cells, beta cells, progenitor cells of pancreatic islet cells, progenitor cells of beta cells, peripheral blood progenitor cells, stem cells or derivatives thereof isolated from adult tissue, retinal progenitor derivative cells, cardiac progenitor derivative cells, osteoprogenitor cells, neural progenitor cells, genetically transformed cells, and any combination thereof. In some cases, the first and second polymer layers are directly sealed along the periphery of the first and second polymer layers. In some cases, the device does not include a support or a frame. In some cases, the first polymer layer, the second polymer layer, or both are non-laminate polymer layers.

[0004] In another embodiment, a method is provided that includes implanting any of the aforementioned devices into a subject having or suspected of having a disease or disorder, the device comprising a plurality of therapeutic cells, thereby treating the disease or disorder. In some cases, the subject is a human. In some cases, the plurality of cells are pancreatic islets or pancreatic islet cells. In some cases, the subject has or is suspected of having diabetes. In some cases, the plurality of cells are allogeneic cells, xenogeneic cells, or autologous cells. In some cases, the device is implanted with the plurality of cells encapsulated within the lumen. In some cases, the device is implanted without the cells encapsulated within the lumen. In some cases, the plurality of cells is introduced into the lumen after the device has been implanted into the subject for a period of time. In some cases, the period of time is at least one week. In some cases, the therapeutic molecule is configured to be released from the device. In some cases, the therapeutic molecule diffuses from the device. In some cases, the therapeutic molecule comprises insulin. In some cases, the implantation is selected from the group consisting of subcutaneous, omental, intraperitoneal, retroperitoneal, and intramuscular implantation. In some cases, the device does not elicit a foreign body response in the subject. In some cases, the therapeutic cells are embedded within the second polymer layer. In some cases, the therapeutic cells adhere to the second polymer layer.

[0005] Incorporation by Reference All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. [Brief description of the drawings]

[0006] Some novel features of the invention are set forth in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings of which:

[0007] [Figure 1A] 1A and 1B show non-limiting examples of cell encapsulation devices according to embodiments of the present disclosure. [Figure 1B] 1A and 1B show non-limiting examples of cell encapsulation devices according to embodiments of the present disclosure.

[0008] [Figure 2A-B] Figures 2A-2E show polycaprolactone scaffolds for cell seeding. Figure 2A shows a polymer scaffold cut into circular wafers. Figure 2B shows the flexibility of the polymer scaffold. Figure 2C shows a highly porous structure within the matrix. Figure 2D shows a scanning electron micrograph of pores within the polymer scaffold. Figure 2E shows a scanning electron micrograph of a cross-section of the polymer scaffold showing the thickness and pores within the matrix. [Figure 2C] Figures 2A-2E show polycaprolactone scaffolds for cell seeding. Figure 2A shows a polymer scaffold cut into circular wafers. Figure 2B shows the flexibility of the polymer scaffold. Figure 2C shows a highly porous structure within the matrix. Figure 2D shows a scanning electron micrograph of pores within the polymer scaffold. Figure 2E shows a scanning electron micrograph of a cross-section of the polymer scaffold showing the thickness and pores within the matrix. [Figure 2D] Figures 2A-2E show polycaprolactone scaffolds for cell seeding. Figure 2A shows a polymer scaffold cut into circular wafers. Figure 2B shows the flexibility of the polymer scaffold. Figure 2C shows a highly porous structure within the matrix. Figure 2D shows a scanning electron micrograph of pores within the polymer scaffold. Figure 2E shows a scanning electron micrograph of a cross-section of the polymer scaffold showing the thickness and pores within the matrix. [Figure 2E] Figures 2A-2E show polycaprolactone scaffolds for cell seeding. Figure 2A shows a polymer scaffold cut into circular wafers. Figure 2B shows the flexibility of the polymer scaffold. Figure 2C shows a highly porous structure within the matrix. Figure 2D shows a scanning electron micrograph of pores within the polymer scaffold. Figure 2E shows a scanning electron micrograph of a cross-section of the polymer scaffold showing the thickness and pores within the matrix.

[0009] [Figure 3A] 3A-3C show another non-limiting example of a cell encapsulation device according to an embodiment of the present disclosure: Fig. 3A shows the macroporous layer of the device, Fig. 3B shows the nanoporous layer of the device, and Fig. 3C shows a cross-sectional view of the device. [Figure 3B] 3A-3C show another non-limiting example of a cell encapsulation device according to an embodiment of the present disclosure: Fig. 3A shows the macroporous layer of the device, Fig. 3B shows the nanoporous layer of the device, and Fig. 3C shows a cross-sectional view of the device. [Figure 3C] 3A-3C show another non-limiting example of a cell encapsulation device according to an embodiment of the present disclosure: Fig. 3A shows the macroporous layer of the device, Fig. 3B shows the nanoporous layer of the device, and Fig. 3C shows a cross-sectional view of the device.

[0010] [Figure 4] FIG. 4 shows a non-limiting example of a method for loading cells into a device according to FIGS. 3A-3C and subsequent placement of the cells in the macroporous layer of the device.

[0011] [Diagram 5] FIG. 5 shows a non-limiting example of subcutaneous placement of a device of the present disclosure into a C57BL / 6 mouse.

[0012] [Figure 6A] FIG. 6A shows a non-limiting example of non-fasting, intraperitoneal glucose tolerance test (ipGTT) results 21 days after implantation of the device. [Figure 6B] FIG. 6B shows a non-limiting example of results from a glucose-stimulated insulin secretion assay.

[0013] [Figure 7]7 shows a cross-sectional view of a device of the present disclosure integrated into a mouse body. Immunofluorescence detection of CD31 and α-smooth muscle actin (α-SMA), as well as DAPI nuclear staining, show that vasculature can grow around and into the device.

[0014] [Figure 8] 8A and 8B show images (at 4x and 10x magnification, respectively) of islets loaded into a device of the present disclosure and implanted subcutaneously in a mouse. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] Disclosed herein is a device for encapsulating biological cells. In some cases, the device can be configured to be implanted into a subject. In some cases, the device of the present disclosure can include a first polymer layer and a second polymer layer. In some cases, the first polymer layer and the second polymer layer can contact at the periphery of the device, thereby forming an enclosed space or lumen between the first polymer layer and the second polymer layer. In some cases, the lumen can be configured to encapsulate a population of biological cells. The device described herein can be designed to contain therapeutic cells (e.g., in the lumen or internal compartment). The device described herein can be designed such that when implanted into a biological subject, the cells contained within the device remain viable and / or functional for a period of time.

[0016] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. It is further noted that the claims may be drafted to exclude any necessary elements. Thus, this description is intended to serve as a premise basis for the use of exclusive terminology such as "solely," "only," and the like in connection with the recitation of claim elements, or the use of a "negative" limitation.

[0017] Particular ranges or numbers are indicated herein by numerical values ​​preceded by the term "about." The term "about" is used herein to mean plus or minus 1%, 2%, 3%, 4%, or 5% of the number to which the term refers.

[0018] As used herein, the terms "subject" and "individual" are used interchangeably and may be any animal, including a mammal (eg, a human or non-human animal).

[0019] As used herein, the terms "treat," "treating," or "treatment" include alleviating, reducing, or ameliorating the symptoms of a disease or condition, preventing additional symptoms, ameliorating or preventing the underlying metabolic cause of the symptoms, inhibiting the disease or condition (e.g., arresting the onset of the disease or condition), relieving the disease or condition, causing regression of the disease or condition, alleviating conditions caused by the disease or condition, or arresting the symptoms of the disease or condition, either prophylactically and / or therapeutically.

[0020] As used herein, the term "connection diameter" refers to the diameter of the open space or channel connecting two adjacent pores. As used herein, the term "pore diameter" refers to the distance between two opposing walls of a pore.

[0021] As used herein, the term "encapsulated" refers to cells that are contained or enclosed within a device of the present disclosure. For example, encapsulated cells may refer to cells that are contained within the lumen or interior space of a device of the present disclosure. In another example, encapsulated cells may refer to cells that are contained within the membrane of the device (e.g., embedded or attached within the macroporous scaffold of the device).

[0022] As used herein, the term "lumen" refers to the internal space or volume of the device of the present disclosure. In some cases, the lumen is created by sealing the periphery of the first polymer layer and the second polymer layer, thereby creating an internal space or volume for encapsulating cells. As used herein, the term "lumen" also encompasses the space in which cells are embedded or attached within the device, for example, the space (e.g., pores, channels, etc.) within the macroporous scaffold of the device.

[0023] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs (e.g., the methods and compositions described herein). Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the methods and compositions described herein, representative exemplary methods and materials are described herein.

[0024] In some embodiments of the present disclosure, the device can include a first polymer layer. In some cases, the first polymer layer can be a polymer membrane or a polymer film. The first polymer layer can include a plurality of pores. In some cases, the first polymer layer is a nanoporous polymer layer.

[0025] In some cases, the plurality of pores present in the first polymer layer can have an average connecting diameter. In some cases, the plurality of pores present in the first polymer layer can have an average connecting diameter in a range of about 10 nm to about 2 μm. For example, the plurality of pores present in the first polymer layer may have an average connection diameter in the range of about 20 nm to about 300 nm, about 30 nm to about 300 nm, about 10 nm to about 200 nm, about 20 nm to about 200 nm, about 30 nm to about 200 nm, about 10 nm to about 100 nm, about 20 nm to about 100 nm, about 30 nm to about 100 nm, about 30 nm to about 50 nm, about 30 nm to about 40 nm, about 20 nm to about 250 nm, about 20 nm to about 150 nm, about 25 to about 150 nm, about 200 nm to about 500 nm, about 350 nm to about 500 nm, about 500 nm to about 1.5 μm, or about 1.0 μm to about 2.0 μm. In some cases, the plurality of pores in the first polymer layer may be about 10 nm, about 20 nm, about 30 nm, about 40 nm, about 50 nm, about 60 nm, about 70 nm, about 80 nm, about 100 nm, about 110 nm, about 120 nm, about 130 nm, about 140 nm, about 150 nm, about 160 nm, about 170 nm, about 180 nm, about 190 nm, about 200 nm, about 210 nm, about 220 nm, about 230 nm, about 240 nm, about 250 nm, about 260 nm, about 270 nm, about 280 nm, about 290 nm, about 300 nm, about 310 nm, about 320 nm, about 330 nm, about 340 nm, about 350 nm, about 360 nm, about 370 nm, about 380 nm, about 390 nm, about 400 nm, about 410 nm, about 420 nm, about 430 nm, about 440 nm, about 450 nm, about 460 nm, about 470 nm, about 480 nm, about 490 nm, about 500 nm, about 510 nm, about 520 nm, about 530 nm, about 540 nm, about 550 nm, about 560 nm, about 570 nm, about 580 nm, about 590 nm, about 600 nm, about 610 nm, about 620 nm, about 630 nm, about 640 nm, about 650 nm, about 660 nm, about 670 nm, about 680 nm, about 690 nm, about 700 nm, about 710 nm, about 720 nm, about 730 The average connection diameter may be about 60 nm, about 370 nm, about 380 nm, about 390 nm, about 400 nm, about 410 nm, about 420 nm, about 430 nm, about 440 nm, about 450 nm, about 460 nm, about 470 nm, about 480 nm, about 490 nm, about 500 nm, about 550 nm, about 600 nm, about 650 nm, about 700 nm, about 750 nm, about 800 nm, about 850 nm, about 900 nm, about 950 nm, about 1.0 μm, about 1.1 μm, about 1.2 μm, about 1.3 μm, about 1.4 μm, about 1.5 μm, about 1.6 μm, about 1.7 μm, about 1.8 μm, about 1.9 μm, or about 2.0 μm. In one embodiment, the average connection diameter may be about 200 nm. In another embodiment, the average connection diameter can be about 2.0 μm.

[0026] In various embodiments, the first polymer layer has a thickness of less than about 2.0 μm, less than about 1.9 μm, less than about 1.8 μm, less than about 1.7 μm, less than about 1.6 μm, less than about 1.5 μm, less than about 1.4 μm, less than about 1.3 μm, less than about 1.2 μm, less than about 1.1 μm, less than about 1.0 μm, less than about 975 nm, less than about 950 nm, less than about 925 nm, less than about 900 nm, less than about 875 nm, less than about 850 nm, less than about 825 nm, less than about 800 nm, less than about 775 nm, less than about 750 nm, less than about 725 nm, less than about 700 nm, less than about 675 nm, less than about 650 nm, less than about 62 The average connection diameter may be less than 5 nm, less than about 600 nm, less than about 575 nm, less than about 550 nm, less than about 525 nm, less than about 500 nm, less than about 475 nm, less than about 450 nm, less than about 425 nm, less than about 400 nm, less than about 375 nm, less than about 350 nm, less than about 325 nm, less than about 300 nm, less than about 275 nm, less than about 250 nm, less than about 225 nm, less than about 200 nm, less than about 175 nm, less than about 150 nm, less than about 125 nm, less than about 100 nm, less than about 75 nm, less than about 50 nm, less than about 25 nm, less than about 20 nm, or less than about 10 nm.

[0027] In some embodiments of the present disclosure, the pores present in the first polymer layer may have an average pore diameter. In some cases, the average pore diameter may range from about 1 μm to about 5 μm. For example, the pores present in the first polymer layer may have an average pore diameter in the range of about 1 μm to about 4.5 μm, about 1 μm to about 4 μm, about 1 μm to about 3.5 μm, about 1 μm to about 3 μm, about 1 μm to about 2.5 μm, about 1 μm to about 2 μm, about 1.5 μm to about 5 μm, about 2 μm to about 5 μm, about 2.5 μm to about 5 μm, about 3 μm to about 5 μm, about 3.5 μm to about 5 μm, or about 4 μm to about 5 μm. In some cases, the plurality of pores present in the first polymer layer may have an average pore diameter of about 1 μm, about 1.5 μm, about 2 μm, about 2.5 μm, about 3 μm, about 3.5 μm, about 4 μm, about 4.5 μm, or about 5 μm. In some cases, the first polymer layer may have an average pore diameter of about 2 μm.

[0028] In various embodiments of the present disclosure, the average pore diameter can be less than about 5 μm, less than about 4.5 μm, less than about 4 μm, less than about 3.5 μm, less than about 3 μm, less than about 2.5 μm, less than about 2 μm, less than about 1.5 μm, or less than about 1 μm.

[0029] In some cases, the plurality of pores present in the first layer may have an average connection diameter that is smaller than the average pore size. In other cases, the plurality of pores present in the first layer may have an average connection diameter that is the same or substantially the same as the average pore size.

[0030] The first polymer layer can have a thickness. In some cases, the first polymer layer can have a thickness of about 200 μm or less, for example, about 200 μm, about 190 μm, about 180 μm, about 170 μm, about 160 μm, about 150 μm, about 140 μm, about 130 μm, about 120 μm, about 110 μm, about 100 μm, about 90 μm, about 80 μm, about 70 μm, about 60 μm, about 50 μm, about 60 μm, about 70 μm, about 80 μm, about 9 ... The first polymer layer may have a thickness of about 10 μm, about 40 μm, about 30 μm, about 29 μm, about 28 μm, about 27 μm, about 26 μm, about 25 μm, about 24 μm, about 23 μm, about 22 μm, about 21 μm, about 20 μm, about 19 μm, about 18 μm, about 17 μm, about 16 μm, about 15 μm, about 14 μm, about 13 μm, about 12 μm, about 11 μm, or about 10 μm. In some cases, the thickness of the first polymer layer may be less than about 10 μm. In some cases, the thickness of the first polymer layer may be greater than about 10 μm but less than about 30 μm. In some cases, the thickness of the first layer may be greater than about 18 μm but less than about 22 μm. In some cases, the thickness of the first polymer layer may be greater than about 30 μm. In some cases, the thickness of the first polymer layer may be about 10 μm to about 200 μm, about 20 μm to about 50 μm, about 50 μm to about 300 μm, or about 100 μm to about 500 μm. In some cases, the thickness of the first polymer layer may be about 10 μm to about 50 μm. In some cases, the thickness of the first polymer layer is about 20 μm.

[0031] The first polymer layer may include a biocompatible polymer. The biocompatible polymer may include, but is not limited to, polycaprolactone (PCL), polytetrafluoroethylene (PTFE), polyethylene (PE), polyvinylidene fluoride (PVDF), methacrylate polymers, polyethyleneimine and dextran sulfate, poly(vinylsiloxane) ecopolymer polyethyleneimine, phosphorylcholine, poly(ethyl methacrylate), polyurethane, poly(ethylene glycol), poly(lactic acid-glycolic acid), hydroxyapatite, poly(lactic acid), polyhydroxyvaleric acid and copolymers, polyhydroxybutyric acid and copolymers, polydioxanone, polyanhydrides, polycyanoacrylates, poly(amino acids), poly(orthoesters), polyesters, silicones, collagen, gelatin, cellulose polymers, chitosan, laminin, and alginate, or combinations thereof. In some cases, the first polymer layer may include polycaprolactone. In some cases, the biocompatible polymer can be biodegradable (e.g., dissolvable in a biological environment). In some cases, the first polymer layer can be manufactured by weaving, casting, extrusion, deposition, emulsion processes, etc. In other cases, the first polymer layer can be manufactured by an electrospinning process or a molding process.

[0032] In some embodiments of the present disclosure, the device can include a second polymer layer. In some cases, the second polymer layer can be a polymer membrane or a polymer film. The second polymer layer can include a plurality of pores. In some cases, the second polymer layer can be a macroporous polymer layer. In some cases, the second polymer layer is a sponge or sponge-like. In some cases, the second polymer layer is a mesh.

[0033] The pores present in the second polymer layer may have an average connection diameter. In some cases, the pores present in the second polymer layer may have an average connection diameter in the range of about 50 μm to about 3 mm. For example, the pores present in the second layer may have an average connection diameter in the range of about 50 μm to about 1 mm, about 200 μm to about 1.5 mm, about 300 μm to about 2 mm, about 400 μm to about 2.5 mm, about 500 μm to about 3 mm, about 100 μm to about 500 μm, about 500 μm to about 1.5 mm, or about 1.5 mm to about 3 mm. In some cases, the plurality of pores present in the second polymer layer may be about 50 μm, about 100 μm, about 150 μm, about 200 μm, about 250 μm, about 300 μm, about 350 μm, about 400 μm, about 450 μm, about 500 μm, about 550 μm, about 600 μm, about 650 μm, about 700 μm, about 750 μm, about 800 μm, about 850 μm, about 900 μm, about 950 μm, , about 1.0 mm, about 1.1 mm, about 1.2 mm, about 1.3 mm, about 1.4 mm, about 1.5 mm, about 1.6 mm, about 1.7 mm, about 1.8 mm, about 1.9 mm, about 2.0 mm, about 2.1 mm, about 2.2 mm, about 2.3 mm, about 2.4 mm, about 2.5 mm, about 2.6 mm, about 2.7 mm, about 2.8 mm, about 2.9 mm, or about 3.0 mm.

[0034] In some cases, the second polymer layer has a thickness of greater than about 50 μm, greater than about 100 μm, greater than about 150 μm, greater than about 200 μm, greater than about 250 μm, greater than about 300 μm, greater than about 350 μm, greater than about 400 μm, greater than about 450 μm, greater than about 500 μm, greater than about 550 μm, greater than about 600 μm, greater than about 650 μm, greater than about 700 μm, greater than about 750 μm, greater than about 800 μm, greater than about 850 μm, greater than about 900 μm, greater than about 950 μm, greater than about 1.0 mm Super, more than about 1.1mm, more than about 1.2mm, more than about 1.3mm, more than about 1.4mm, more than about 1.5mm, more than about 1.6mm, more than about 1.7mm, more than about 1.8mm, more than about 1.9mm, more than about 2.0mm, about 2.1mm The average connection diameter may be greater than about 2.2 mm, greater than about 2.3 mm, greater than about 2.4 mm, greater than about 2.5 mm, greater than about 2.6 mm, greater than about 2.7 mm, greater than about 2.8 mm, greater than about 2.9 mm, or greater than about 3.0 mm.

[0035] The pores present in the second polymer layer may have an average pore diameter. In some cases, the pores present in the second polymer layer may have an average pore diameter in the range of about 100 μm to about 3 mm. For example, the pores present in the second layer may have an average pore diameter in the range of about 100 μm to about 1 mm, about 200 μm to about 1.5 mm, about 300 μm to about 2 mm, about 400 μm to about 2.5 mm, about 500 μm to about 3 mm, about 100 μm to about 500 μm, about 500 μm to about 1.5 mm, or about 1.5 mm to about 3 mm. In some cases, the plurality of pores present in the second polymer layer may have a diameter of about 100 μm, about 150 μm, about 200 μm, about 250 μm, about 300 μm, about 350 μm, about 400 μm, about 450 μm, about 500 μm, about 550 μm, about 600 μm, about 650 μm, about 700 μm, about 750 μm, about 800 μm, about 850 μm, about 900 μm, about 950 μm, ...0 μm, about 1500 μm, about 1000 μm, about 1500 μm, about 1500 μm, about 1500 μm, about 1500 μm, about 1500 μm, about 1500 μm, about 1500 μm, about 1500 μm, about 1500 μm, about 1500 μm, about 1500 μm, about 1500 μm, about 1500 μm, about 1500 μm, about 1500 μm, about 1500 μm, about 1500 μm, about 1500 μm, about 1500 μm, about 15 The plurality of pores present in the second polymer layer may have an average pore diameter of about 0.0 mm, about 1.1 mm, about 1.2 mm, about 1.3 mm, about 1.4 mm, about 1.5 mm, about 1.6 mm, about 1.7 mm, about 1.8 mm, about 1.9 mm, about 2.0 mm, about 2.1 mm, about 2.2 mm, about 2.3 mm, about 2.4 mm, about 2.5 mm, about 2.6 mm, about 2.7 mm, about 2.8 mm, about 2.9 mm, or about 3.0 mm. The plurality of pores present in the second polymer layer may be stochastically organized.

[0036] In some cases, the second polymer layer has a thickness of greater than about 100 μm, greater than about 150 μm, greater than about 200 μm, greater than about 250 μm, greater than about 300 μm, greater than about 350 μm, greater than about 400 μm, greater than about 450 μm, greater than about 500 μm, greater than about 550 μm, greater than about 600 μm, greater than about 650 μm, greater than about 700 μm, greater than about 750 μm, greater than about 800 μm, greater than about 850 μm, greater than about 900 μm, greater than about 950 μm, greater than about 1.0 mm, greater than about More than 1.1mm, more than about 1.2mm, more than about 1.3mm, more than about 1.4mm, more than about 1.5mm, more than about 1.6mm, more than about 1.7mm, more than about 1.8mm, more than about 1.9mm, more than about 2.0mm, more than about 2.1mm, It can have an average pore diameter of greater than about 2.2 mm, greater than about 2.3 mm, greater than about 2.4 mm, greater than about 2.5 mm, greater than about 2.6 mm, greater than about 2.7 mm, greater than about 2.8 mm, greater than about 2.9 mm, or greater than about 3.0 mm.

[0037] The second polymer layer may have a thickness. In some cases, the second polymer layer may have a thickness of about 200 μm to about 1 mm. For example, the second polymer layer may have a thickness of about 200 μm to about 500 μm, about 250 μm to about 450 μm, about 300 μm to about 500 μm, about 350 μm to about 600 μm, about 400 μm to about 800 μm, about 550 μm to about 950 μm, or about 500 μm to about 1 mm. In some cases, the second polymer layer can have a thickness of about 200 μm, about 250 μm, about 300 μm, about 350 μm, about 400 μm, about 450 μm, about 500 μm, about 550 μm, about 600 μm, about 650 μm, about 700 μm, about 750 μm, about 800 μm, about 850 μm, about 900 μm, about 950 μm, or about 1 mm. In some cases, the second polymer layer can have a thickness of about 500 μm.

[0038] In some cases, the second polymer layer may have a thickness of about 1 mm to about 5 mm. For example, the second polymer layer may have a thickness of about 1 mm to about 5 mm, about 2 mm to about 4 mm, about 3 mm to about 5 mm, about 1 mm to about 3 mm, about 2.5 mm to about 5 mm, or about 1.5 mm to about 4.5 mm. In some cases, the second polymer layer may have a thickness of about 1 mm, about 1.1 mm, about 1.2 mm, about 1.3 mm, about 1.4 mm, about 1.5 mm, about 1.6 mm, about 1.7 mm, about 1.8 mm, about 1.9 mm, about 2.0 mm, about 2.1 mm, about 2.2 mm, about 2.3 mm, about 2.4 mm, about 2.5 mm, about 2.6 mm, about 2.7 mm, about 2.8 mm, about 2.9 mm, The second polymer layer may have a thickness of about 3.0 mm, about 3.1 mm, about 3.2 mm, about 3.3 mm, about 3.4 mm, about 3.5 mm, about 3.6 mm, about 3.7 mm, about 3.8 mm, about 3.9 mm, about 4.0 mm, about 4.1 mm, about 4.2 mm, about 4.3 mm, about 4.4 mm, about 4.5 mm, about 4.6 mm, about 4.7 mm, about 4.8 mm, about 4.9 mm, or about 5.0 mm. In some cases, the second polymer layer may have a thickness of about 2 mm.

[0039] The second polymer layer can have a pore density of about 0.01 g / cm. 3 ~about 0.1g / cm3 For example, the second polymer layer may have a pore density of about 0.01 g / cm 3 , about 0.02g / cm 3 , about 0.03g / cm 3 , about 0.04g / cm 3 , about 0.05g / cm 3 , about 0.06g / cm 3 , about 0.07g / cm 3 , about 0.08g / cm 3 , about 0.09g / cm 3 , or about 0.1 g / cm 3 The pore density may be

[0040] The second polymer layer may comprise a biocompatible polymer. Biocompatible polymers include, but are not limited to, polycaprolactone (PCL), polytetrafluoroethylene (PTFE), polyethylene (PE), polyvinylidene fluoride (PVDF), methacrylate polymers, polyethyleneimine and dextran sulfate, poly(vinylsiloxane) ecopolymer polyethyleneimine, phosphorylcholine, poly(ethyl methacrylate), polyurethane, poly(ethylene glycol), poly(lactic acid-glycolic acid), hydroxyapatite, poly(lactic acid), polyhydroxyvaleric acid and copolymers, polyhydroxybutyric acid and copolymers, polydioxanone, polyanhydrides, polycyanoacrylates, poly(amino acids), poly(orthoesters), polyesters, collagen, gelatin, cellulose polymers, chitosan, laminin, and alginate, or combinations thereof. In some cases, the second polymer layer may comprise polycaprolactone (PCL). In some cases, the biocompatible polymer can be biodegradable (e.g., dissolvable in a biological environment). In some cases, the second polymer layer can be manufactured by weaving, extrusion, casting, deposition, emulsion processes, etc. In other cases, the second polymer layer can be manufactured by an electrospinning process or a molding process.

[0041] In some embodiments, the second polymer layer can act as a scaffold for biological cells in the lumen of the device. For example, biological cells contained in the lumen of the device can attach to and / or be embedded in the surface of the pores or channels of the second polymer layer. In some cases, the surface of the second polymer layer can be coated with one or more agents (e.g., growth factors) that promote the survival and / or growth and / or protection of biological cells in the device. In some cases, the second polymer layer can promote the vascularization of cells encapsulated in the device. For example, the second polymer layer can allow blood vessels to grow up to or penetrate the second polymer layer so that they are in close proximity to the biological cells encapsulated in the device. The blood vessels penetrating the second polymer layer can provide a supply of oxygen and / or nutrients to the cells encapsulated in the device, promoting the viability of the cells in the device.

[0042] In some cases, the device of the present disclosure may increase the viability of encapsulated cells compared to transplanted naked nucleus cells (e.g., cells that are not encapsulated in the device). The device described herein may be particularly well suited for encapsulating cells that prefer to attach to or be embedded in a substrate, for example, cells that have a preferred three-dimensional structure or arrangement. In certain instances, the macroporous scaffold of the device provides a substrate for cells to be embedded or attached thereon. In some cases, encapsulated cells embedded or attached in the macroporous scaffold may increase viability compared to encapsulated cells that are not embedded or attached to the scaffold (e.g., suspended in the lumen of the device).

[0043] In various embodiments, the first polymer layer and the second polymer layer can each be a non-laminated polymer layer. In other words, each of the first polymer layer and the second polymer layer can consist of a single polymer layer.

[0044] In some embodiments, the device of the present disclosure may further comprise a third polymer layer. In some cases, the third polymer layer may comprise a plurality of pores. In some cases, the plurality of pores may have the average connection diameter and / or average pore diameter of those disclosed for the first polymer layer. In some cases, the third polymer layer is a nanoporous polymer layer. In some cases, the first and third polymer layers have the same or similar average connection diameter and / or average pore diameter. In some cases, the first and third polymer layers have different average connection diameter and / or average pore diameter. In some cases, the third polymer layer may be composed of a biocompatible polymer, such as any biocompatible polymer disclosed for the first polymer layer. In some cases, the first and third polymer layers are composed of the same biocompatible polymer material. In some cases, the first and third polymer layers are composed of different biocompatible polymer materials.

[0045] In such devices that include a third polymer layer, the third polymer layer may contact the first polymer layer at the periphery of the device, or the third polymer layer may contact the first and second polymer layers at the periphery of the device, thereby forming an enclosed space or lumen between the first and third polymer layers. In such cases, the second polymer layer may be enclosed within the device (e.g., within the lumen, or lining the third polymer layer).

[0046] In various embodiments, the devices of the present disclosure may have an overall thickness of about 200 μm to about 5 mm. For example, the devices of the present disclosure may have an overall thickness of about 200 μm, about 250 μm, about 300 μm, about 350 μm, about 400 μm, about 450 μm, about 500 μm, about 550 μm, about 600 μm, about 650 μm, about 700 μm, about 750 μm, about 800 μm, about 850 μm, about 900 μm, about 950 μm, about 1 mm, about 1.1 mm, about 1.2 mm, about 1.3 mm, about 1.4 mm, about 1.5 mm, about 1.6 mm, about 1.7 mm, about 1.8 mm, about 1.9 mm, about 2.0 mm, about 2.1 mm, about 2.2 mm, about 2.3 mm, about 2.4 mm, about 2.5 mm, about 2.6 mm, about 2.7 mm, about 2.8 mm, about 2.9 mm, about 3.0 mm, about 3.1 mm, about 3.2 mm, about 3.3 mm, about 3.4 mm, about 3.5 mm, about 3.6 mm, about 3.7 mm, about 3.8 mm, about 3.9 mm, about 3.1 mm, about 3.2 ...3 mm, about 3.4 mm, about 3.5 mm, about 3.6 mm, about 3.7 mm, about 3.8 mm, about 3.9 mm, about 3.1 mm, about 3.2 In some embodiments, the thickness of the insulating layer may be about 2.0 mm, about 2.3 mm, about 2.4 mm, about 2.5 mm, about 2.6 mm, about 2.7 mm, about 2.8 mm, about 2.9 mm, about 3.0 mm, about 3.1 mm, about 3.2 mm, about 3.3 mm, about 3.4 mm, about 3.5 mm, about 3.6 mm, about 3.7 mm, about 3.8 mm, about 3.9 mm, about 4.0 mm, about 4.1 mm, about 4.2 mm, about 4.3 mm, about 4.4 mm, about 4.5 mm, about 4.6 mm, about 4.7 mm, about 4.8 mm, about 4.9 mm, or about 5.0 mm.

[0047] In some cases, the devices of the present disclosure are about 1 cm 2 ~Approx. 100cm 2 For example, the devices of the present disclosure may have a surface area of ​​about 1 cm 2 , about 5cm 2 , about 10cm 2 , about 15cm 2 , about 20cm 2 , about 25cm 2 , about 30cm 2 , about 35cm 2 , about 40cm 2 , about 45cm 2 , about 50cm 2 , approx. 55cm 2 , about 60cm 2 , approx. 65cm 2 , about 70cm 2 , about 75cm 2 , about 80cm 2 , approx. 85cm 2 , about 90cm 2 , about 95cm 2 , or about 100 cm 2 may have a surface area of

[0048] In some cases, the device provided herein may have a two-dimensional planar shape.For example, the two-dimensional planar shape may be, but is not limited to, a circle, an ellipse, a square, or a rectangle.In some cases, the device provided herein may have a three-dimensional shape.For example, the three-dimensional shape may be, but is not limited to, a sphere, a cone, a cube, a cylinder, a triangle, a hexagon, a tetrahedron, a pyramid, or an octagon.

[0049] In some cases, the devices provided herein may be used for transplantation of various different cell types into biological subjects. In some cases, the biological cells may be eukaryotic cells. In some cases, the biological cells may be prokaryotic cells. In general, the biological cells may be derived from any source. In some cases, the cells may be bacterial cells, yeast cells, insect cells, or fungal cells. In some cases, the cells may be mammalian cells. Non-limiting examples of mammalian cell types that may be used include thyroid cells, parathyroid cells, bone marrow cells, mesenchymal stem cells, stromal cells, pluripotent stem cells, induced pluripotent stem cells, embryonic stem cells, vascular cells, cells derived from adipose tissue, cells derived from bone marrow, intestinal cells or cells derived therefrom, pancreatic islets or islet cells, Sertoli cells, beta cells, precursor cells of pancreatic islet cells, precursor cells of beta cells, peripheral blood precursor cells, stem cells or derivatives thereof isolated from adult tissues, retinal progenitor derived cells, cardiac progenitor derived cells, osteoprogenitor cells, neural progenitor cells, genetically transformed cells, or any combination thereof. In some cases, the cell may be a pancreatic islet or a pancreatic islet cell. In some cases, the cell may be a beta cell or a beta-like cell. In some cases, the cell may be a stem cell. The derived cell may include an isolated subset of primary tissue cells in an unaltered state or in a state modified by culture conditions or by the introduction of materials into the cell. In some cases, the cell may be an allogeneic cell. In some cases, the cell may be an autologous cell. In some cases, the cell may be a xenogeneic cell. In some cases, the cell may be a cell that has been engineered to express one or more recombinant proteins.

[0050] In some cases, the cells may be derived from a human. In other cases, the cells may be derived from a non-human animal, including, but not limited to, a non-human primate, a livestock animal, a domestic pet (e.g., dog, cat, sheep, cow, horse) or a laboratory animal. For example, the non-human animal may be an ape (e.g., chimpanzee, baboon, gorilla, or orangutan), an Old World monkey (e.g., rhesus monkey), a New World monkey, a dog, a cat, a bison, a camel, a cow, a deer, a pig, a donkey, a horse, a mule, a llama, a sheep, a goat, a buffalo, a reindeer, a yak, a mouse, a rat, a rabbit, a whale, or other non-human animal. The encapsulated cells may be derived from the subject (e.g., autologous cells), another donor (e.g., allogeneic cells), or another species (e.g., xenogeneic cells). In a non-limiting example, the cells may be xenogeneic islets or islet cells (e.g., from a pig) that are transplanted into a human subject.

[0051] The cells can be introduced into the lumen of the device, and the device can be immediately (within a day) implanted into a subject. Alternatively, the cells can be introduced into the lumen of the device, and the device containing the encapsulated cells therein can be cultured, for example, for more than a day, before implantation to allow the cells to grow and / or become embedded into the scaffold of the device. In some cases, the cells can be cultured for up to 12 hours, up to 1 day, up to 2 days, up to 3 days, up to 4 days, up to 5 days, up to 6 days, up to 7 days, or more than 7 days before the device is implanted into a subject.

[0052] In some cases, the number of cells introduced into the lumen of the device is at least 100 cells, at least 200 cells, at least 300 cells, at least 400 cells, at least 500 cells, at least 1000 cells, at least 2000 cells, at least 3000 cells, at least 4000 cells, at least 5000 cells, at least 10,000 cells, at least 20,000 cells, at least 30,000 cells, at least 40,000 cells, at least 50,000 cells, at least 60,000 cells, at least 70,000 cells, at least 80,000 cells, at least 90,000 cells, at least 100,000 cells, at least 15 ... cells, at least 100,000 cells, at least 200,000 cells, at least 300,000 cells, at least 400,000 cells, at least 500,000 cells, at least 600,000 cells, at least 700,000 cells, at least 800,000 cells, at least 900,000 cells, at least 1 million, at least 5 million, at least 10 million, at least 50 million, at least 100 million, at least 200 million, at least 300 million, at least 400 million, at least 500 million, at least 600 million, at least 700 million, at least 800 million, at least 900 million, or at least 1 billion. The number of cells introduced into the lumen of the device can vary and can be empirically determined.

[0053] In some embodiments, the cells introduced into the lumen of the device may be modified. The modification may include genetic modification or epigenetic modification. The genetic modification may include insertion, deletion, mutation, or inversion. The modification may be introduced into the cells through gene editing, for example, using the CRISPR / Cas gene editing system. The cells may be modified to alter (e.g., increase, decrease, or eliminate) the expression of a protein of interest. The cells may be modified to increase the expression of a protein of interest. The genetic modification to alter the expression of a protein of interest may include modifications to regulatory regions involved in the expression of the protein of interest. Regulatory regions may include, but are not limited to, promoters, transcription factors, enhancers, silencers, 5'UTRs, and 3'UTRs. The cells may be modified to generate structural or functional variants of the protein of interest. The cells introduced into the lumen may be recombinant cells.

[0054] In some aspects, the cells may be therapeutic cells, e.g., cells that produce and release or secrete therapeutic molecules that provide a therapeutic benefit to the biological subject. In general, the device may be configured such that the therapeutic molecule may be released from the device to achieve, e.g., a therapeutic response in the biological subject. The therapeutic molecule may be a protein. In some cases, the protein may be a naturally occurring protein. In some cases, the protein may be a recombinant protein. The protein may be a hormone or an enzyme. The protein may be insulin or an analog thereof, erythropoietin, a coagulation modulator (e.g., a coagulant or anticoagulant), a thrombolytic enzyme, alpha-galactosidase A, deoxyribonuclease I, glucocerebrosidase, idronidase, N-acetylgalactosamine-4-sulfatase, a growth hormone or an analog thereof, a gonadotropin, calcitonin, glucagon, interferon, interleukin, thrombopoietin, estrogen, testosterone, or granulocyte colony-stimulating factor. The coagulation agent may be factor VII. The therapeutic molecule may be factor A, factor VIII, or factor IX. The gonadotropin may be follitropin, chorionic gonadotropin, follicle-stimulating hormone, or luteinizing hormone. The interferon may be interferon alpha, interferon beta, or interferon gamma. The interleukin may be IL-1, IL-2, IL-3, IL-4, IL-10, IL-11, IL-13, IL-15, IL-17, or IL-18. In some cases, the therapeutic molecule is an antibody. In one non-limiting example, the cell is a pancreatic islet or islet cell, and the therapeutic molecule is insulin (e.g., to treat a diabetic subject). The type of cells used in the device is generally determined based on the desired therapeutic molecule and the therapeutic response desired in the subject.

[0055] In some aspects, the device provided herein can be configured to be implanted into a subject. In some cases, the device can be used to implant or transfer cells into a subject. In some cases, the device can be implanted into a subject by subcutaneous implantation, intramuscular implantation, subdermal implantation, intraperitoneal implantation, retroperitoneal implantation, omental implantation, brain implantation, subface implantation, bone implantation, eye implantation, organ implantation, adjacent to an organ (such as liver). The site of implantation can be selected based on the diseased / damaged tissue that needs treatment. For the treatment of diseases such as diabetes mellitus (DM), the device can be placed in a clinically convenient site such as subcutaneous space or omentum.

[0056] In certain embodiments, the device may be completely sealed along the edges of the device, thereby forming a completely enclosed internal space or lumen. In other embodiments, the device may be open at one or more locations at the end of the device, allowing access to the internal space or lumen of the device. When the device includes two openings to the lumen of the device, the two openings may be located opposite each other, such as, for example, opposite sides of a planar device. In some cases, the device of the present disclosure does not include a support or frame. For example, the first polymer layer and the second polymer layer may be placed on top of each other and sealed along the periphery of the layers. In some cases, the device of the present disclosure may have a support or frame. In such embodiments, the device may be flexible or may have a flexible structure such that it can be rolled up. In some cases, the device may be folded or rolled up before implantation, and then unfolded or unrolled after implantation.

[0057] In some aspects, cells can be loaded into the lumen of the device before implantation into a subject. In some cases, after the cells are loaded into the lumen of the device, the cells can be embedded in the macroporous scaffold of the second polymer layer (Figure 4). In other embodiments, the device (without any cells loaded) can be implanted into a subject. Then, after the device is implanted for a period of time, the device can be loaded with cells. For example, the device can be implanted into a subject at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 1 week, at least 1.5 weeks, at least 2 weeks, at least 2.5 weeks, at least 3 weeks, at least 3.5 weeks, or at least 4 weeks before loading the cells into the device. In some aspects, the device can further include a port for loading the cells into the device. In some cases, the port is a tube for introducing the cells into the lumen of the device. In certain embodiments, the tube can be attached to the device by sealing the first and second layers to the outer wall of the tube at the opening. In certain embodiments, the first end of the tube may be placed in the opening and within the lumen such that a minimum volume of the lumen is occupied by the tube while allowing the lumen of the device to be filled with fluid. The second end of the tube, distal to the first end, may be used as a port for introducing fluid into the lumen by the tube. The length of the tube may be selected based on several factors. For example, if a population of cells is to be loaded into the lumen of the device ex vivo, e.g., prior to placing the device in a subject. If the tube is to be used to introduce fluid into and / or remove fluid from the lumen of the device after placement of the device in a subject, a longer tube may be used. In some cases, the port may be accessible after the device is implanted in a subject, such that the device may be pre-implanted without any cells in the lumen, and the lumen may be loaded with cells at a period of time after implantation. In some aspects, the device may be pre-loaded with support cells or factors (e.g., growth factors) prior to the introduction of therapeutic cells.

[0058] The devices herein may be designed such that therapeutic molecules (e.g., those produced by and secreted from therapeutic cells) may diffuse out of the lumen or internal compartment of the device. Additionally, the devices may allow molecules present on the outside of the device to penetrate into the lumen or internal compartment of the device. In some cases, the devices may allow oxygen to diffuse into and / or out of the device. In some aspects, the devices may promote blood vessel growth in or around the implantation site of the device (e.g., promote vascularization of the device).

[0059] When cell-laden implantable devices are isolated from the immune system by being encased in a cell-impermeable layer, the implantable device often stimulates a local inflammatory response, termed foreign body transport (FBR), which has long been recognized as limiting the functionality of implanted devices that require the transport of solutes. The FBR is well described in the literature and is composed of three main layers. The innermost FBR layer adjacent to the implanted device is composed of macrophages and foreign body giant cells. These cells form a monolayer of closely opposed cells on the surface of the implanted device. The middle FBR layer, distal to the first layer with respect to the device, is a large area (30-100 microns) composed primarily of fibroblasts and fibrous matrix. The outermost FBR layer is a loose connective granulation tissue that contains new blood vessels. Upon introduction of the FBR, the implanted device is isolated from the in vivo environment limiting the exchange of molecules with the implanted device, limiting the usefulness of the implanted device, as well as leading to the death of any cells provided within the implanted device. In some cases, the devices of the present disclosure may cause no foreign body reaction, no substantial foreign body reaction, or a limited foreign body reaction, such that cells encapsulated within the device remain viable for a period of time. In some cases, the foreign body reaction may be evidenced by fibrosis around the implanted device. In some cases, the devices of the present disclosure may cause no fibrosis, no substantial fibrosis, or limited fibrosis around the implanted device. Figures 8A and 8B illustrate devices of the present disclosure demonstrating a lack of fibrosis around the implanted device.

[0060] In some aspects, the devices disclosed herein, upon implantation into a subject, may support the viability of cells present within the lumen of the device for at least 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 2 years, at least 3 years, at least 4 years, at least 5 years, or more.

[0061] In some aspects, the devices disclosed herein may be substantially impermeable to cells present outside the device such that the cells present outside the device do not enter the device. For example, in some cases, the devices are substantially impermeable to immune cells such that the immune cells present outside the device cannot enter the lumen of the device. In some aspects, the devices disclosed herein may be substantially impermeable to molecules present outside the device such that the molecules do not enter the device. For example, in some cases, the device is substantially impermeable to immunoglobulins such that the concentration of any immunoglobulins accessing the lumen of the device is lower than the level required for an immune response to the cells present within the lumen of the device. In another example, the devices of the present disclosure may substantially restrict cytokines from entering the lumen of the device such that the concentration of cytokines accessing the lumen of the device is lower than the level required for an immune response to the cells present within the lumen of the device. In some aspects, the devices are configured to facilitate the exchange of nutrients with the encapsulated cells and / or the diffusion of metabolic waste products from the device. In some cases, the devices provided herein are configured to allow the passage of small molecules (including therapeutic molecules) but substantially inhibit the passage of large molecules.

[0062] In some aspects, the exterior surface of the device can be modified by coating with one or more agents that improve the function of the device. For example, molecules that promote vascularization of the device or inhibit immune or inflammatory responses to the device can be placed on the exterior of the device. In some cases, agents can be placed within the device (e.g., loaded into the lumen of the device or coated on the inner surface of the device). In some cases, agents can diffuse from the lumen of the device and act on the external environment. Additionally or alternatively, agents within the lumen of the device can act on cells placed within the lumen of the device (e.g., maturation factors that act on partially mature stem cells). Such molecules or agents may include, but are not limited to, VEGF (vascular endothelial growth factor), PDGF (platelet-derived growth factor), FGF-1 (fibroblast growth factor), angiopoietins MCP-1, αvβ3, αvβ5, CD-31, VE-cadherin, ephrin, plasminogen activator, angiogenin, Del-1, aFGF (acidic fibroblast growth factor), vFGF (basic fibroblast growth factor), follistatin, G-CSF (granulocyte colony stimulating factor), HGF (hepatocyte growth factor), leptin, placental growth factor, PD-ECGF (platelet-derived endothelial growth factor), and the like.

[0063] Further provided herein is a method for using the subject device. In one aspect, the device of the present disclosure can be implanted into a subject. In some cases, the device can be implanted into a subject with the device's lumen preloaded with cells. In other cases, the device can be implanted into a subject without the device's lumen preloaded with cells. In such cases, the device can be loaded with cells after a period of time (e.g., by using a pre-implantation port or a tube of the device). In some cases, the method can include using the subject device to implant into a subject cells that produce and secrete therapeutic molecules. In some cases, the therapeutic molecules can diffuse from the device into the subject's environment, and the therapeutic molecules can have a therapeutic effect on the subject.

[0064] In some cases, the method can be used to treat the disease or disorder of the subject.The disease or disorder can be characterized by the lack of functional cells or the insufficient production of protein.In some cases, the disease or disorder can be, but is not limited to, diabetes, anemia, coagulation disorder (e.g., hemophilia), cardiovascular disease, Fabry disease, Gaucher disease, mucopolysaccharidosis, growth hormone deficiency, infertility, osteoporosis, hypoglycemia, Parkinson's disease, or muscular dystrophy.

[0065] In a non-limiting example, the method can be used to transplant pancreatic islets or islet cells into a subject, where the islets or islet cells produce and secrete insulin. In such cases, the method can be used to treat a subject who has or is suspected of having diabetes. The diabetes can be type 1 diabetes. The device can include islet cells or can include stem cells that can differentiate into islet cells. In certain embodiments, pluripotent stem cells (PSCs) can be differentiated into islet cells within the device, and then the device containing the differentiated islet cells is placed in the subject (e.g., in the omentum adjacent to the pancreas or liver). In some cases, the device can include PSCs, and the device can be implanted adjacent to the pancreas or liver of the subject. EXAMPLES

[0066] Example 1. Cell encapsulation device.

[0067] 1A and 1B illustrate non-limiting examples of cell encapsulation devices according to embodiments of the present disclosure. FIG. 1A illustrates a first polymer layer. The first polymer layer includes a plurality of pores having an average connection diameter of about 200 nm and an average pore diameter of about 2 μm. FIG. 1B illustrates a second polymer layer. The second polymer layer includes a plurality of pores having an average pore diameter of about 500 μm to about 3 mm and an average connection diameter of about 100 μm to about 500 μm.

[0068] Example 2. Polymer scaffolds.

[0069] A three-dimensional polymer scaffold was designed to facilitate transplantation of pancreatic islets in a manner that allows optimal packing of the islets with better spatial distribution. The scaffold was designed with a stochastic macroporous structure that served to allow the islets to be embedded within it as well as to allow ingrowth of blood vessels to vascularize the islets (Figures 2A-2E).

[0070] Example 3. Cell encapsulation device.

[0071] The device was constructed with nanoporous and macroporous layers constructed of polycaprolactone (e.g., scaffold according to Example 2) (Figures 3A-3C). The nanoporous membrane was about 20 μm thick, had a connection diameter of about 200 nm, and consisted of pores with a pore diameter of about 2 μm. The macroporous membrane was about 500 μm thick, and consisted of macroscopic pores with connection diameters and pore diameters of 200-400 μm. To create the device, the macroporous and nanoporous membranes were overlapped and directly sealed along the perimeter.

[0072] Example 4. Syngeneic islet transplantation into diabetic mice.

[0073] The devices were loaded with approximately 500 syngeneic C57BL / 6 islets, sealed, and subcutaneously implanted into C57BL / 6 mice as described in Example 3 (Figure 5). An intraperitoneal glucose tolerance test (IPGTT) was performed on day 21. Glucose tolerance tests measure the clearance from the body of a glucose load injected into the peritoneal cavity and can be used to detect disorders of glucose metabolism that may be related to human conditions such as diabetes or metabolic syndrome. Before administering a glucose solution via intraperitoneal (IP) injection, the animals were fasted for approximately 16 hours and fasting blood glucose levels were measured. Blood glucose levels were then measured at various time points over the next 2 hours.

[0074] IPGTT was performed on day 21 with 500 islets encapsulated within the subcutaneously implanted device and 500 islets implanted directly into the kidney capsule. The islets within the device followed the same glucose curve as the islets within the kidney capsule and showed a similar glucose response (Figure 6A).

[0075] Furthermore, insulin secretion was analyzed using a glucose-stimulated insulin secretion assay. The glucose stimulation index is a metric for quantifying beta cell function by comparing the ratio of insulin release in high glucose conditions relative to resting conditions. The ratio of insulin secreted in high glucose conditions to low glucose conditions was used to calculate the glucose stimulation index. As illustrated in Figure 6B, islets encapsulated in a device as described in Example 3 had a greater glucose stimulation index than islets directly transplanted into the kidney capsule (KC).

[0076] Example 5. Evaluation of the device after engraftment.

[0077] Devices were implanted subcutaneously for >30 days to allow for engraftment. After sacrifice, tissues were excised and hyperfixed using Z-Fix at 4°C for 24 hours, then washed with PBS. Tissues and devices were imaged using CLARITY (Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging / Immunostaining / in situ-hybridization-compatible Tissue Hydrogel), a technology that embeds tissue in a hydrogel matrix. The lipids were made transparent to create an optically clear hybrid that transmits light and is transparent to molecules. Markers included were DAPI for nuclear staining, as well as CD31 and α-smooth muscle actin (α-SMA) to stain blood vessels. Figure 7 illustrates staining of blood vessels infiltrated within and around the device.

[0078] Figures 8A and 8B illustrate immunohistochemical staining of sections of the device after engraftment in mice, illustrating pancreatic islets embedded within the macroporous scaffold layer of the device (arrows).

[0079] While preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be used to practice the invention. It is intended that the following claims define the scope of the invention, and that methods and structures within the scope of these claims and their equivalents be covered thereby. The present invention provides, for example, the following items. (Item 1) a) a first polymer layer comprising a plurality of pores having an average connecting diameter of less than about 500 nm; b) a second polymer layer comprising a plurality of pores having an average connection diameter of greater than about 100 μm; A device comprising: The first polymer layer and the second polymer layer define a lumen for enclosing a plurality of cells. (Item 2) 2. The device of claim 1, wherein the plurality of pores present in the first polymer layer have an average connection diameter of about 10 nm to about 200 nm. (Item 3) 2. The device of claim 1, wherein the plurality of pores present in the first polymer layer have an average connection diameter of about 180 nm to about 220 nm. (Item 4) 4. The device according to any one of items 1 to 3, wherein the plurality of pores present in the second polymer layer have an average connection diameter of about 100 μm to about 500 μm. (Item 5) 4. The device according to any one of items 1 to 3, wherein the plurality of pores present in the second polymer layer have an average connection diameter of about 200 μm to about 400 μm. (Item 6) 5. The device according to any one of items 1 to 4, wherein the plurality of pores present in the first polymer layer have an average pore diameter of about 1 μm to about 5 μm. (Item 7) 7. The device according to any one of items 1 to 6, wherein the plurality of pores present in the second polymer layer have an average pore diameter of about 500 μm to about 3 mm. (Item 8) 8. The device according to any one of items 1 to 7, wherein the plurality of pores present in the second polymer layer have an average pore diameter of about 100 μm to about 1 mm. (Item 9) 9. The device of any one of items 1 to 8, wherein the first polymer layer has a thickness of about 10 μm to about 200 μm. (Item 10) 10. The device of any one of items 1 to 9, wherein the first polymer layer has a thickness of about 20 μm to about 50 μm. (Item 11) 10. The device of any one of items 1 to 9, wherein the first polymer layer has a thickness of about 18 μm to about 22 μm. (Item 12) 11. The device of any one of items 1 to 10, wherein the second polymer layer has a thickness of about 1 mm to about 5 mm. (Item 13) 11. The device of any one of items 1 to 10, wherein the second polymer layer has a thickness of about 400 μm to about 600 μm. (Item 14) 13. The device of any one of items 1 to 12, further comprising a third polymer layer having an average connection diameter of less than about 500 nm. (Item 15) 15. The device according to any one of items 1 to 14, further comprising a port for introducing the plurality of cells into the lumen of the device. (Item 16) 16. The device of any one of items 1-15, wherein the first polymer layer, the second polymer layer, the third polymer layer, or a combination thereof comprises a biocompatible polymer selected from the group consisting of polycaprolactone (PCL), polyvinylidene fluoride (PVDF), polytetrafluoroethylene (PTFE), polyethylene (PE), methacrylate polymers, polyethyleneimine, polyethyleneimine-dextran sulfate, poly(vinylsiloxane) ecopolymer polyethyleneimine, phosphorylcholine, poly(ethyl methacrylate), polyurethane, poly(ethylene glycol) (PEG), poly(lactic-glycolic acid) (PLGA), hydroxyapatite, poly(lactic acid), polyhydroxyvaleric acid and copolymers thereof, polyhydroxybutyric acid and copolymers thereof, polydioxanone, polyanhydrides, polycyanoacrylates, poly(amino acids), poly(orthoesters), polyesters, collagen, gelatin, cellulose polymers, chitosan, alginates, laminins, and any combination thereof. (Item 17) 17. The device of claim 16, wherein the biocompatible polymer is polycaprolactone. (Item 18) 18. The device of any one of items 1-17, wherein the plurality of cells is selected from the group consisting of thyroid cells, parathyroid cells, bone marrow cells, mesenchymal stem cells, stromal cells, pluripotent stem cells, induced pluripotent stem cells, embryonic stem cells, vascular cells, cells derived from adipose tissue, cells derived from bone marrow, intestinal cells or cells derived therefrom, pancreatic islets or islet cells, Sertoli cells, beta cells, progenitor cells of pancreatic islet cells, progenitor cells of beta cells, peripheral blood progenitor cells, stem cells or derivatives thereof isolated from adult tissue, retinal progenitor derived cells, cardiac progenitor derived cells, osteoprogenitor cells, neural progenitor cells, genetically transformed cells, and any combination thereof. (Item 19) 19. The device of any one of items 1 to 18, wherein the first polymer layer and the second polymer layer are directly sealed along a perimeter of the first polymer layer and the second polymer layer. (Item 20) 20. The device according to any one of items 1 to 19, wherein the device does not include a support or a frame. (Item 21) 21. The device of any one of items 1 to 20, wherein the first polymer layer, the second polymer layer, or both are non-laminated polymer layers. (Item 22) 22. A method comprising the step of implanting the device according to any one of items 1 to 21 into a subject having or suspected of having a disease or disorder, wherein the device comprises a plurality of therapeutic cells, thereby treating the disease or disorder. (Item 23) 23. The method of claim 22, wherein the subject is a human. (Item 24) 24. The method of claim 22 or 23, wherein the plurality of cells is a pancreatic islet or pancreatic islet cells. (Item 25) 25. The method according to any one of items 22 to 24, wherein the subject has or is suspected of having diabetes. (Item 26) 26. The method according to any one of items 22 to 25, wherein the plurality of cells are allogeneic cells, xenogeneic cells, or autologous cells. (Item 27) 27. The method of any one of items 22 to 26, wherein the device is implanted with a plurality of cells encapsulated within the lumen. (Item 28) 28. The method of any one of items 22-27, wherein the device is implanted without cells encapsulated within the lumen. (Item 29) 30. The method of claim 28, wherein a plurality of cells are introduced into the lumen after the device has been implanted in the subject for a period of time. (Item 30) 30. The method of claim 29, wherein the period of time is at least one week. (Item 31) 31. The method according to any one of items 22 to 30, wherein the therapeutic molecule is configured to be released from the device. (Item 32) 32. The method of claim 31, wherein the therapeutic molecule diffuses out of the device. (Item 33) 33. The method of claim 31 or 32, wherein the therapeutic molecule comprises insulin. (Item 34) 34. The method according to any one of items 22 to 33, wherein the implantation is selected from the group consisting of subcutaneous, omental, intraperitoneal, retroperitoneal, and intramuscular implantation. (Item 35) 35. The method of any one of items 22 to 34, wherein the device does not induce a foreign body reaction in the subject. (Item 36) 36. The method of any one of items 22 to 35, wherein the therapeutic cells are embedded within the second polymer layer. (Item 37) 37. The method of any one of items 22 to 36, wherein the therapeutic cells are attached to the second polymer layer.

Claims

[Claim 1] The invention described in this specification.

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