Cell packaging device and use method thereof

Abstract

Disclosed herein are devices for encapsulating biological cells and suitable for implantation in a subject. In various aspects of the present disclosure, the device may include a plurality of polymer layers. In one aspect, a device includes a first polymer layer and a second polymer layer. In some cases, the first polymer layer may be a nanoporous polymer layer. In some cases, the second polymer layer may be a macroporous polymer layer. The first polymer layer and the second polymer layer may define a lumen for encapsulating a population of cells. The device may be used to transplant cells that produce a therapeutic agent into a subject (e.g., in order to treat a disease).

Description

[0001] This application is a divisional application of the Chinese patent application with the application date of March 11, 2020, application number 202080020730.1, and invention name “Cell encapsulation device and method of use thereof” (the application date of the corresponding PCT application is March 11, 2020, and the application number is PCT / US2020 / 022063).

[0002] Cross-references

[0003] This application claims the benefit of U.S. Provisional Application No. 62 / 816,675, filed on March 11, 2019, which is incorporated herein by reference in its entirety. Background Art

[0004] Transplanting cells into a subject to produce therapeutic molecules can be an interesting alternative to the use of exogenous drugs. Encapsulation of cell transplants can, for example, prevent the proliferation of transplanted cells in the host and can enhance the viability of transplanted cells. The present disclosure provides methods and devices for improving transplantation into a subject, including improving the viability and function of transplanted cells. Summary of the invention

[0005] In one aspect, a device is provided, comprising: a) a first polymer layer comprising a plurality of pores having an average connectivity diameter of less than about 500 nm; and b) a second polymer layer comprising a plurality of pores having an average connectivity diameter of greater 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 connectivity 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 connectivity 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 connectivity 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 connectivity 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 holes present in the second polymer layer have an average pore diameter of about 500 μm to about 3 mm. In some cases, the plurality of holes present in the second polymer layer have 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 includes a third polymer layer having an average connectivity diameter of less than about 500 nm. In some cases, the device further includes a port for introducing a plurality of cells into the lumen of the device.In some cases, the first polymer layer, the second polymer layer, 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)ecopolymere polyethyleneimine, phosphorylcholine, poly(ethyl methacrylate), polyurethane, poly(ethylene glycol) (PEG), poly(lactic-co-glycolic acid) (PLGA), hydroxyapatite, poly(lactic acid), polyhydroxyvalerate and copolymers thereof, polyhydroxybutyrate and copolymers thereof, 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 are selected from: 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 from intestinal cells, islets of Langerhans or islet cells, Sertoli cells, β cells, islet progenitor cells, β progenitor cells, peripheral blood progenitor cells, stem cells or derivatives thereof separated from adult tissue, retinal progenitor cell-derived cells, cardiac progenitor cell-derived cells, osteoprogenitor cells, neuronal progenitor cells, genetically transformed cells, and any combination thereof. In some cases, the first polymer layer and the second polymer layer are directly sealed along the periphery of the first polymer layer and the second polymer layer. 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-laminated polymer layers.

[0006] On the other hand, a method is provided, comprising: implanting a device according to any of the foregoing situations into a subject suffering from or suspected of having a disease or condition, wherein the device comprises a plurality of therapeutic cells, thereby treating the disease or condition. In some cases, the subject is a human being. In some cases, the plurality of cells are pancreatic islets or pancreatic islet cells. In some cases, the subject suffers from 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 together with a plurality of cells encapsulated in the lumen. In some cases, the device is implanted without any cells being encapsulated in the lumen. In some cases, after the device is implanted in the subject for a period of time, a plurality of cells are introduced into the lumen. In some cases, the period of time is at least 1 week. In some cases, the therapeutic molecule is configured to be released from the device. In some cases, the therapeutic molecule diffuses out from the device. In some cases, the therapeutic molecule comprises insulin. In some cases, the implantation is selected from the group consisting of: subcutaneous implantation, omental implantation, intraperitoneal implantation, retroperitoneal implantation, and intramuscular implantation. In some cases, the device does not induce a foreign body reaction in the subject. In some cases, the therapeutic cells are embedded within the second polymer layer. In some cases, the therapeutic cells are attached to the second polymer layer.

[0007] Incorporation by reference

[0008] 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

[0009] Some of the novel features of the invention are set forth in the appended claims. A better understanding of the features and advantages of the invention will be obtained by referring to the following detailed description and accompanying drawings which set forth an illustrative embodiment in which the principles of the invention are employed, wherein:

[0010] Figure 1A and Figure 1B Depicted are non-limiting examples of cell encapsulation devices according to embodiments of the present disclosure.

[0011] Figure 2A-2E Depicts the polycaprolactone scaffold used for cell seeding. Figure 2A Depicts a polymer scaffold cut into circular slices. Figure 2B Delineating the flexibility of polymer scaffolds. Figure 2C Depicting the highly porous architecture within the matrix. Figure 2D Scanning electron micrograph depicting pores within a polymer scaffold. Figure 2E Scanning electron microscope image depicting a cross-section of a polymer scaffold, showing the thickness and pores within the matrix.

[0012] Figure 3A-3C Another non-limiting example of a cell encapsulation device according to an embodiment of the present disclosure is depicted. Figure 3A Depicts the macroporous layer of the device. Figure 3B Depicts the nanoporous layer of the device. Figure 3C A cross-sectional view of the device is shown.

[0013] Figure 4 Figure 2. Cells are loaded into Figures 3A-3C A non-limiting example of a method in a device is shown, and the subsequent arrangement of cells into the macroporous layer of the device.

[0014] Figure 5 A non-limiting example of subcutaneous placement of a device of the present disclosure into a C57BL / 6 mouse is depicted.

[0015] Fig. 6A Shown are non-limiting examples of results of a non-fasting intraperitoneal glucose tolerance test (ipGTT) on day 21 after device implantation. Figure 6B Non-limiting examples of results of a glucose stimulated insulin secretion test are depicted.

[0016] Figure 7 A cross-sectional view depicting a device of the present disclosure integrated into a mouse. Immunofluorescence detection of CD31 and alpha smooth muscle actin (α-SMA) as well as DAPI nuclear staining demonstrated that vasculature could grow around and within the device.

[0017] Fig. 8A and Figure 8B Images depicting pancreatic islets loaded into a device of the present disclosure and implanted subcutaneously in a mouse (4X and 10X magnification, respectively). DETAILED DESCRIPTION

[0018] Disclosed herein is a device for encapsulating biological cells. In some cases, the device can be configured to be implanted in a subject. In some cases, the device of the present disclosure may include a first polymer layer and a second polymer layer. In some cases, the first polymer layer and the second polymer layer may contact at the periphery of the device, thereby forming an encapsulated space or lumen between the first polymer layer and the second polymer layer. In some cases, the lumen is configured to encapsulate a biological cell group. The device described herein can be designed to accommodate therapeutic cells (e.g., in a lumen or an internal compartment). The device described herein can be designed so that when the device is transplanted into a biological subject, the cells contained in the device remain alive and / or functional for a period of time.

[0019] As used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It should be further noted that a claim may be drafted to exclude any optional elements. Thus, this statement is intended to serve as antecedent basis for using exclusive terminology such as "only," "only," and the like in connection with the recitation of claim elements or for using a "negative" limitation.

[0020] Certain ranges or numbers are expressed herein with the term "about" preceding the numerical value. 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.

[0021] As used herein, the terms "subject" and "individual" are used interchangeably and can be any animal, including mammals (eg, a human or non-human animal).

[0022] As used herein, the terms "treat," "treating," or "treatment" include alleviating, alleviating, or ameliorating symptoms of a disease or condition, preventing additional symptoms, ameliorating or preventing potential metabolic causes of symptoms, inhibiting the disease or condition, e.g., arresting the progression of the disease or condition, slowing the disease or condition, causing regression of the disease or condition, relieving the condition caused by the disease or condition, or prophylactically and / or therapeutically terminating symptoms of the disease or condition.

[0023] The term "connectivity diameter" as used herein means the diameter of an open space or channel connecting two adjacent pores. The term "pore diameter" as used herein means the distance between two opposing walls of a pore.

[0024] The term "encapsulated" as used herein means that a cell is contained or encapsulated within a device of the present disclosure. For example, an encapsulated cell may refer to a cell contained within a lumen or interior space of a device of the present disclosure. In another example, an encapsulated cell may refer to a cell contained within a membrane of the device (e.g., embedded within or attached to a macroporous scaffold of the device).

[0025] The term "lumen" as used herein 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 to form an internal space or volume to encapsulate cells. The term "lumen" as used herein 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.

[0026] 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 the present disclosure (e.g., the methods and compositions described herein) belongs. Representative illustrative methods and materials are now described, but any methods and materials similar or equivalent to those described herein can also be used to implement or test the methods and compositions described herein.

[0027] In some aspects of the present disclosure, the device may include a first polymer layer. In some cases, the first polymer layer may be a polymer film or a polymer film. The first polymer layer may include a plurality of pores. In some cases, the first polymer layer is a nanoporous polymer layer.

[0028] In some cases, the multiple holes present in the first polymer layer can have an average connectivity diameter. In some cases, the multiple holes present in the first polymer layer can have an average connectivity diameter of about 10nm to about 2 μm. For example, the multiple holes present in the first polymer layer can have an average connectivity diameter of about 20nm to about 300nm, about 30nm to about 300nm, about 10nm to about 200nm, about 20nm to about 200nm, about 30nm to about 200nm, about 10nm to about 100nm, about 20nm to about 100nm, about 30nm to about 100nm, about 30nm to about 50nm, about 30nm to about 40nm, about 20nm to about 250nm, about 20nm to about 150nm, about 25 to about 150nm, about 200nm to about 500nm, about 350nm to about 500nm, about 500nm to about 1.5 μm, or an average connectivity diameter of about 1.0 μm to about 2.0 μm. In some cases, the plurality of pores in the first polymer layer can have a diameter of 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 In one aspect, the average connectivity diameter can be about 200 nm. In another aspect, the average connectivity diameter can be about 2.0 μm.

[0029] In various aspects, the first polymer layer can have 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 In some embodiments, the present invention relates to an average connectivity diameter of less than about 625 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.

[0030] In some aspects of the present disclosure, the plurality of holes present in the first polymer layer may have an average pore diameter. In some cases, the average pore diameter may be in the range of about 1 μm to about 5 μm. For example, the plurality of holes present in the first polymer layer may have an average pore diameter 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 holes 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 can have an average pore diameter of about 2 μm.

[0031] In various aspects of the present disclosure, the average pore diameter may 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.

[0032] In some cases, the plurality of pores present in the first layer may have an average connected 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 connected diameter that is the same or substantially the same as the average pore size.

[0033] The first polymer layer may have a thickness. In some cases, the first polymer layer may 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 8 ... In some cases, the thickness of the first polymer layer may be less than about 10 μm. In some cases, 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 can 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 can be about 10 μm to about 50 μm. In some cases, the thickness of the first polymer layer is about 20 μm.

[0034] The first polymer layer can include biocompatible polymers.Biocompatible polymers can include, without limitation, polycaprolactone (PCL), polytetrafluoroethylene (PTFE), polyethylene (PE), polyvinylidene fluoride (PVDF), methacrylate polymers, polyethyleneimine and dextran sulfate, poly (vinylsiloxane) ecopolymere polyethyleneimine, phosphorylcholine, poly (ethyl methacrylate), polyurethane, poly (ethylene glycol), poly (lactic acid-glycolic acid), hydroxyapatite, poly (lactic acid), polyhydroxyvalerate and copolymer, polyhydroxybutyrate and copolymer, polydioxanone, polyanhydride, polycyanoacrylate, poly (amino acid), poly (orthoester), polyester, silicone, collagen, gelatin, cellulose polymer, chitosan, laminin and alginate, or their combination.In some cases, the first polymer layer can include polycaprolactone.In some cases, biocompatible polymers can be biodegradable (for example, soluble in biological environment). In some cases, the first polymer layer can be prepared by weaving, by casting, by extrusion, by deposition, by an emulsification process, etc. In other cases, the first polymer layer can be prepared by an electrospinning process or a template process.

[0035] In some aspects of the present disclosure, the device may include a second polymer layer. In some cases, the second polymer layer may be a polymer film or a polymer film. The second polymer layer may include a plurality of pores. In some cases, the second polymer layer may 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.

[0036] The multiple holes present in the second polymer layer can have an average connectivity diameter. In some cases, the multiple holes present in the second polymer layer can have an average connectivity diameter of about 50 μm to about 3 mm. For example, the multiple holes present in the second layer can have an average connectivity diameter 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 an average connectivity diameter of 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 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 10 ... , 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 in average connectivity diameter.

[0037] In some cases, the second polymer layer can have 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, greater than about 1.1 mm, greater than about 1.2 mm, greater than about 1.3 mm, greater than about 1.4 mm, greater than about 1.5 mm, greater than about 1.6 mm, greater than about 1.7 mm, greater than about 1.8 mm, greater than about 1.9 mm, greater than about 2.0 mm, greater than about 2.1 mm, 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 in average connectivity diameter.

[0038] The plurality of holes present in the second polymer layer can have an average pore diameter. In some cases, the plurality of holes present in the second polymer layer can have an average pore diameter of about 100 μm to about 3 mm. For example, the plurality of holes present in the second layer can have an average pore diameter 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 an average pore diameter of 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, about 10 ... The plurality of pores present in the second polymer layer may be randomly organized.

[0039] In some cases, the second polymer layer can have 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, or greater than about 1.0 mm. An average pore diameter of greater than about 1.1 mm, greater than about 1.2 mm, greater than about 1.3 mm, greater than about 1.4 mm, greater than about 1.5 mm, greater than about 1.6 mm, greater than about 1.7 mm, greater than about 1.8 mm, greater than about 1.9 mm, greater than about 2.0 mm, greater than about 2.1 mm, 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.

[0040] The second polymer layer may have a thickness. In some cases, the second polymer layer has 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, 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.

[0041] In some cases, the second polymer layer can have a thickness of about 1 mm to about 5 mm. For example, the second polymer layer can 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 can 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 In some cases, the second polymer layer can have a thickness of about 2 mm. In some cases, the second polymer layer can 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 can have a thickness of about 2 mm.

[0042] The second polymer layer can have a pore density. In some cases, the second polymer layer can have a pore density of about 0.01 g / cm 3 To about 0.1g / cm 3 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 0.1g / cm 3 The hole density.

[0043] The second polymer layer can include biocompatible polymers.Biocompatible polymers can include, but are not limited to, polycaprolactone (PCL), polytetrafluoroethylene (PTFE), polyethylene (PE), polyvinylidene fluoride (PVDF), methacrylate polymers, polyethyleneimine and dextran sulfate, poly (vinylsiloxane) ecopolymere polyethyleneimine, phosphorylcholine, poly (ethyl methacrylate), polyurethane, poly (ethylene glycol), poly (lactic acid-glycolic acid), hydroxyapatite, poly (lactic acid), polyhydroxyvalerate and copolymer, polyhydroxybutyrate and copolymer, polydioxanone, polyanhydride, polycyanoacrylate, poly (amino acid), poly (orthoester), polyester, collagen, gelatin, cellulose polymer, chitosan, laminin and alginate, or their combination.In some cases, the second polymer layer can include polycaprolactone (PCL).In some cases, biocompatible polymers can be biodegradable (for example, soluble in biological environment). In some cases, the second polymer layer can be prepared by weaving, by extrusion, by casting, by deposition, by an emulsification process, etc. In other cases, the second polymer layer can be prepared by an electrospinning process or a template process.

[0044] In some respects, the second polymer layer can serve as the support of the biological cells in the lumen of the device.For example, the biological cells contained in the lumen of the device can be attached to the surface of the second polymer layer and / or be embedded in the hole or channel of the second polymer layer.In some cases, the surface of the second polymer layer can be coated with one or more medicaments (such as growth factors), and the one or more medicaments promote the survival and / or growth and / or protection of the biological cells in the device.In some cases, the second polymer layer can promote the vascularization of the cell encapsulated in the device.For example, the second polymer layer can allow blood vessels to grow into the second polymer layer or penetrate the second polymer layer, so that blood vessels are closely approached to the biological cells encapsulated in the device.The blood vessels penetrating the second polymer layer can provide the supply of oxygen and / or nutrients to the cells encapsulated in the device and can promote the viability of the cells in the device.

[0045] In some cases, the device of the present disclosure can enhance the viability of encapsulated cells relative to naked cells (e.g., cells not encapsulated in the device) of transplantation. The device described herein can be particularly well suited for encapsulating cells (e.g., cells with a preferred three-dimensional architecture or arrangement) that are preferably attached to or embedded in a substrate. In a specific embodiment, the macroporous support of the device provides a substrate so that cells are embedded or attached thereto. In some cases, encapsulated cells embedded in or attached to a macroporous support can have enhanced viability compared to encapsulated cells that are not embedded or attached to a support (e.g., freely floating in the lumen of a device).

[0046] In various aspects, the first polymer layer and the second polymer layer can each be a non-laminated polymer layer. In other words, the first polymer layer and the second polymer layer can each be composed of a single polymer layer.

[0047] In some aspects, the device of the present disclosure may further include a third polymer layer. In some cases, the third polymer layer may include a plurality of holes. In some cases, a plurality of holes may have those average connectivity diameters and / or average pore diameters disclosed for the first polymer layer. In some cases, the third polymer layer is a nanoporous polymer layer. In some cases, the first polymer layer and the third polymer layer have the same or similar average connectivity diameters and / or average pore diameters. In some cases, the first polymer layer and the third polymer layer have different average connectivity diameters and / or average pore diameters. In some cases, the third polymer layer may be composed of a biocompatible polymer, for example, any biocompatible polymer disclosed for the first polymer layer. In some cases, the first polymer layer and the third polymer layer are composed of the same biocompatible polymer material. In some cases, the first polymer layer and the third polymer layer are composed of different biocompatible polymer materials.

[0048] In such a device including a third polymer layer, the third polymer layer may be in contact with the first polymer layer at the periphery of the device, or the third polymer layer may be in contact with the first polymer layer and the second polymer layer at the periphery of the device, thereby forming an enclosed space or lumen between the first polymer layer and the third polymer layer. In this case, the second polymer layer may be enclosed within the device (e.g., within the lumen or as a lining of the third polymer layer).

[0049] In various aspects, the devices of the present disclosure can have an overall thickness of about 200 μm to about 5 mm. For example, the devices of the present disclosure can 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 4.1 mm, about 4.2 mm, about 4.3 mm, about 4.4 mm, about A total thickness of 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 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.

[0050] In some cases, the disclosed devices may have a diameter of about 1 cm 2 To about 100cm 2 For example, the device of the present disclosure may have a surface area of ​​about 1 cm 2 、About 5cm 2 , about 10cm 2 、Approx. 15cm 2 、About 20cm 2 、Approx. 25cm 2 、About 30cm 2 、Approx. 35cm 2 、Approx. 40cm 2 、Approx. 45cm 2 、Approx. 50cm 2 、Approx. 55cm 2 、Approx. 60cm 2 、Approx. 65cm 2 、Approx. 70cm 2 、Approx. 75cm 2 、Approx. 80cm 2 、Approx. 85cm 2 、Approx. 90cm 2 、Approx. 95cm 2 Or about 100cm 2 surface area.

[0051] In some cases, the devices provided herein can have a two-dimensional planar shape. For example, the two-dimensional planar shape can be, without limitation, circular, elliptical, square, or rectangular. In some cases, the devices provided herein can have a three-dimensional shape. For example, the three-dimensional shape can be, without limitation, spherical, conical, cubic, cylindrical, triangular, hexagonal, tetrahedral, pyramidal, or octagonal.

[0052] In some cases, the device provided herein can be used to transplant various cell types into biological subjects. In some cases, biological cells can be eukaryotic cells. In some cases, biological cells can be prokaryotic cells. In general, biological cells can be derived from any source. In some cases, cells can be bacterial cells, yeast cells, insect cells or fungal cells. In some cases, cells can be mammalian cells. Non-limiting examples of mammalian cell types that can 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 from intestinal cells, islets or islet cells, Sertoli cells, β cells, islet progenitor cells, β progenitor cells, peripheral blood progenitor cells, stem cells or derivatives thereof separated from adult tissues, retinal progenitor cell derived cells, cardiac progenitor cell derived cells, osteoprogenitor cells, neuronal progenitor cells, genetically transformed cells, or any combination thereof. In some cases, cells can be islets or islet cells. In some cases, the cell can be a beta cell or a beta-like cell. In some cases, the cell can be a stem cell. The derived cell can include a subpopulation of primary tissue cells that are in an unchanged state, or modified by culture conditions or by introducing materials into the cell. In some cases, the cell can be an allogeneic cell. In some cases, the cell can be an autologous cell. In some cases, the cell can be a xenogeneic cell. In some cases, the cell can be a cell that has been engineered to express one or more recombinant proteins.

[0053] In some cases, the cell can be derived from humans. In other cases, the cell can be derived from non-human animals, including but not limited to, non-human primates, livestock animals, domestic pets (e.g., dogs, cats, sheep, cattle, horses) or laboratory animals. For example, non-human animals can be apes (e.g., chimpanzees, baboons, gorillas or orangutans), old world monkeys (e.g., rhesus monkeys), new world monkeys, dogs, cats, bison, camels, cattle, deer, pigs, donkeys, horses, mules, alpacas, sheep, goats, buffaloes, reindeers, yaks, mice, rats, rabbits, whales or any other non-human animals. Encapsulated cells can be from subjects (e.g., autologous cells), from another donor (e.g., allogeneic cells) or from other species (e.g., xenogeneic cells). In non-limiting examples, cells can be xenogeneic (e.g., from pigs) islets or islet cells transplanted into human subjects.

[0054] Cells can be introduced into the lumen of the device and the device can be implanted in the subject immediately (in one day). Alternatively, cells can be introduced into the lumen of the device and the device accommodating the cells encapsulated therein can be cultured within a period of time (e.g., greater than one day) to allow cells to proliferate and / or be embedded in the support of the device before implantation. In some cases, before the device is implanted in the subject, cells can be cultured 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 greater than 7 days.

[0055] 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, ... In some embodiments, the number of cells introduced into the lumen of the device can vary and can be determined empirically.

[0056] In some embodiments, the cells introduced into the lumen of the device can be modified. Modifications may include genetic modifications or epigenetic modifications. Genetic modifications may include insertions, deletions, mutations or inversions. Modifications may be introduced into cells by gene editing, for example, by using a CRISPR / Cas gene editing system. Cells may be modified to change (e.g., increase, decrease or eliminate) the expression of a protein of interest. Cells may be modified to increase the expression of a protein of interest. Genetic modifications that change the expression of a protein of interest may include modifications to regulatory regions involved in the expression of a protein of interest. Regulatory regions may include, but are not limited to, promoters, transcription factors, enhancers, silencers, 5'UTRs and 3'UTRs. Cells may be modified to produce structural or functional variants of a protein of interest. Cells introduced into the lumen may be recombinant cells.

[0057] In some aspects, the cell can be a therapeutic cell, for example, a cell that produces and releases or secretes a therapeutic molecule, which provides a therapeutic benefit to a biological subject. In general, the device can be configured so that the therapeutic molecule can be released from the device, so as to achieve a therapeutic response, for example, in a biological subject. The therapeutic molecule can be a protein. In some cases, the protein can be a naturally occurring protein. In some cases, the protein can be a recombinant protein. The protein can be a hormone or an enzyme. The protein can be insulin or its analogs, erythropoietin, a coagulation regulator (for example, a coagulant or an anticoagulant), a thrombolytic enzyme, alpha galactosidase A, deoxyribonuclease I, glucocerebrosidase, iduronidase, N-acetylgalactosamine-4-sulfatase, growth hormone or its analogs, gonadotropin, calcitonin, glucagon, interferon, interleukin, thrombopoietin, estrogen, testosterone or granulocyte colony stimulating factor. The coagulant can be factor VII A, factor VIII or factor IX. Gonadotropin can be follicle stimulating hormone, chorionic gonadotropin, follicle stimulating hormone or luteinizing hormone. Interferon can be interferon-stimulating, interferon-stem or interferon-activator. Interleukin can 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, therapeutic molecule is an antibody. In a non-limiting example, the cell is an islet of Langerhans or an islet cell, and the therapeutic molecule is insulin (for example, to treat diabetic subjects). The cell type used in the device is generally determined based on the desired therapeutic molecule and the desired therapeutic response in the subject.

[0058] In some respects, device provided herein can be configured to be implanted in a subject. In some cases, device can be used for cell transplantation or transfer to a subject. In some cases, device can be implanted by subcutaneous implantation, intramuscular implantation, subdermal implantation, intraperitoneal implantation, retroperitoneal implantation, omentum implantation, implantation in the brain, implantation under the face, implantation in the bone, eye implantation, implantation in an organ, adjacent organs (e.g., liver, etc.) implantation and implanted in a subject. The implanted position can be selected based on the diseased tissue / damaged tissue that needs to be treated. In order to treat diseases such as diabetes (DM), device can be placed in a clinically convenient position such as subcutaneous space or omentum.

[0059] In certain embodiments, device can be completely sealed along the edge of the device, thereby forming a completely encapsulated interior space or lumen. In other embodiments, device can be opened at one or more positions at the edge of the device to allow entry into the interior space or lumen of the device. When device includes two openings for entering the lumen of the device, two openings can be positioned relative to each other, such as, on the opposite sides of a planar device. In some cases, device of the present disclosure does not include a support or framework. For example, the first polymer layer and the second polymer layer can be laid on each other and sealed along the periphery of the layer. In some cases, device of the present disclosure can have a support or framework. In such embodiments, device can be flexible or have a flexible structure so that it can be wound. In some cases, device can be folded or wound before implantation, and then unfolded or spread out after implantation.

[0060] In some aspects, cells can be loaded into the lumen of the device prior to implantation in a subject. In some cases, after the cells have been loaded into the lumen of the device, the cells can be embedded within the macroporous scaffold of the second polymer layer ( Figure 4). In other aspects, the device (wherein no cells are loaded) can be implanted in a subject. After the device has been implanted for a period of time, the device can be subsequently loaded with cells. For example, the device can be implanted in 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 the device is loaded with cells. In some aspects, the device can further include a port so that cells are loaded into the device. In some cases, the port is a pipeline for introducing cells into the lumen of the device. In certain embodiments, the pipeline can be attached to the device by sealing the first and second layers to the outer wall of the pipeline at the opening. In certain embodiments, the first end of the pipeline can be placed at the opening and positioned in the lumen so that the minimum volume of the lumen is occupied by the pipeline while allowing the fluid to be loaded into the lumen of the device. The second end of the pipeline at the far side of the first end can be used as a port so that the fluid is introduced into the lumen via the pipeline. The length of the pipeline can be selected based on a variety of factors. For example, when the cell colony is loaded into the lumen of the device in vitro, for example, before the device is placed in the experimenter, a shorter pipeline can be used. If the pipeline will be used to introduce fluid into the lumen of the device and / or remove fluid from the lumen of the device after the device is placed in the experimenter, a longer pipeline can be used. In some cases, after the device has been implanted in the experimenter, the port can be accessible so that the device can be pre-implanted and no cell is accommodated in the lumen, and the lumen can be loaded with cells when implanting a period of time later. In some aspects, the device can be pre-loaded with supporting cells or factors (for example, growth factors) before the introduction of therapeutic cells.

[0061] The device herein can be designed so that therapeutic molecules (e.g., those produced or secreted by therapeutic cells) can diffuse out of the lumen or internal compartment of the device. In addition, the device can allow molecules present outside the device to penetrate into the lumen or internal compartment of the device. In some cases, the device can allow oxygen to diffuse into and / or diffuse out of the device. In some aspects, the device can promote blood vessel growth into or around the implantation site of the device (e.g., promote vascularization of the device).

[0062] When the implantable device that holds cells is enclosed in the cell-impermeable layer and isolated relative to the immune system, the implantable device often stimulates a local inflammatory reaction called foreign body reaction (FBR), which has long been considered to limit the function of the implanted device that needs solute transport. FBR has been fully described in the literature and is composed of three main layers. The innermost layer of the FBR adjacent to the implanted device is composed of macrophages and foreign body giant cells. These cells form a single layer of cells that are closely relative to the surface of the implanted device. The FBR middle layer located at the far side of the first layer relative to the device is a wider area (30-100 microns) mainly composed of fibroblasts and fibrous matrix. The outermost layer of the FBR is a loose granular connective tissue comprising new blood vessels. Once FBR is induced, the implanted device is isolated from the internal environment, thereby limiting the molecular exchange with the implanted device, and also limiting the effectiveness of the implanted device, causing the death of any cell provided in the implanted device. In some cases, the devices of the present disclosure may not provoke a foreign body response, may not provoke a significant foreign body response, or may provoke a limited foreign body response such that cells encapsulated within the device remain viable for a period of time. In some cases, the foreign body response may be evidenced by fibrosis around the implanted device. In some cases, the devices of the present disclosure may not provoke fibrosis around the implanted device, may not provoke significant fibrosis, or may provoke limited fibrosis. Fig. 8A and Figure 8B Depicting a device of the present disclosure showing the absence of fibrosis surrounding the implanted device.

[0063] In some aspects, a device disclosed herein, once implanted into a subject, can support the survival of cells present in the lumen of the device for at least one week, two weeks, three weeks, four weeks, one 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 longer.

[0064] In some aspects, the device disclosed herein can be substantially impermeable to cells present outside the device, so that cells present outside the device cannot enter the device. For example, in some cases, the device is substantially impermeable to immune cells, so that immune cells present outside the device cannot enter the lumen of the device. In some aspects, the device disclosed herein can be substantially impermeable to molecules present outside the device, so that the molecules cannot enter the device. For example, in some cases, the device is substantially impermeable to immunoglobulins, so that the concentration of any immunoglobulin entering the lumen of the device is lower than the level required for the immune response of cells present in the lumen of the device. In another example, the device disclosed herein can substantially limit cytokines from entering the lumen of the device, so that the concentration of cytokines entering the lumen of the device is lower than the level required for the immune response of cells present in the lumen of the device. In some aspects, the device is configured to promote nutrient exchange with encapsulated cells and / or promote the diffusion of metabolic waste to the outside of 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.

[0065] In some aspects, the outer 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 disposed on the outside of the device. In some cases, the agent can be disposed within the device (e.g., loaded within the lumen of the device or coated on the inner surface of the device). In some cases, the agent can diffuse out of the lumen of the device and act on the external environment. Additionally or alternatively, the agent within the lumen of the device can act on cells disposed within the lumen of the device (e.g., a maturation factor that acts 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), angiopoietin MCP-1, α v β3, α v β5, CD-31, VE-cadherin, ephrin, plasminogen activator, angiopoietin, 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 cell growth factor), etc.

[0066] Further provided herein is a method for using a subject device. On the one hand, the device of the present disclosure can be implanted in a subject. In some cases, the device can be implanted in a subject when cells are preloaded into the lumen of the device. In other cases, the device can be implanted in a subject when cells are not preloaded into the lumen of the device. In such cases, the device can be loaded with cells after a period of time (for example, by using a pre-implanted port or the pipeline of the device). In some cases, the method can include using the subject device to transplant cells that produce and secrete therapeutic molecules into a subject. In some cases, therapeutic molecules can diffuse from the device and enter the environment of the subject so that the therapeutic molecules exert a therapeutic effect on the subject.

[0067] In some cases, the method can be used to treat a disease or condition of a subject. A disease or condition can be a disease or condition characterized by lack of functional cells or insufficient protein production. In some cases, a disease or condition can be non-limitingly diabetes, anemia, coagulation disorders (e.g., hemophilia), cardiovascular disease, Fabry disease, Gaucher disease, mucopolysaccharide metabolism disease, growth hormone deficiency, infertility, osteoporosis, hypoglycemia, Parkinson's disease, or muscular dystrophy.

[0068] In non-limiting examples, the method can be used for transplanting islets or islet cells into a subject, wherein the islets or islet cells produce and secrete insulin. In such cases, the method can be used for treating a subject suffering from or suspected of having diabetes. 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 (PSC) can differentiate into islet cells in the device, and the device for accommodating differentiated islet cells is then placed in a subject (e.g., in omentum, adjacent to pancreas or liver). In some cases, the device can include PSC and the device can be implanted near the pancreas or liver of a subject.

[0069] The present invention provides embodiments including but not limited to the following:

[0070] 1. A device comprising:

[0071] a) a first polymer layer comprising a plurality of pores having an average connected diameter of less than about 500 nm;

[0072] b) a second polymer layer comprising a plurality of pores having an average connected diameter greater than about 100 μm;

[0073] The first polymer layer and the second polymer layer define a lumen for encapsulating a plurality of cells.

[0074] 2. The device of embodiment 1, wherein the plurality of pores present in the first polymer layer have an average connectivity diameter of about 10 nm to about 200 nm.

[0075] 3. The device of embodiment 1, wherein the plurality of pores present in the first polymer layer have an average connected diameter of about 180 nm to about 220 nm.

[0076] 4. The device of any one of embodiments 1-3, wherein the plurality of pores present in the second polymer layer have an average connected diameter of about 100 μm to about 500 μm.

[0077] 5. The device of any one of embodiments 1-3, wherein the plurality of pores present in the second polymer layer have an average connected diameter of about 200 μm to about 400 μm.

[0078] 6. The device of any one of embodiments 1-4, wherein the plurality of pores present in the first polymer layer have an average pore diameter of about 1 μm to about 5 μm.

[0079] 7. The device of any one of embodiments 1-6, wherein the plurality of pores present in the second polymer layer have an average pore diameter of about 500 μm to about 3 mm.

[0080] 8. The device of any one of embodiments 1-7, wherein the plurality of pores present in the second polymer layer have an average pore diameter of about 100 μm to about 1 mm.

[0081] 9. The device of any one of embodiments 1-8, wherein the first polymer layer has a thickness of about 10 μm to about 200 μm.

[0082] 10. The device of any one of embodiments 1-9, wherein the first polymer layer has a thickness of about 20 μm to about 50 μm.

[0083] 11. The device of any one of embodiments 1-9, wherein the first polymer layer has a thickness of about 18 μm to about 22 μm.

[0084] 12. The device of any one of embodiments 1-10, wherein the second polymer layer has a thickness of about 1 mm to about 5 mm.

[0085] 13. The device of any one of embodiments 1-10, wherein the second polymer layer has a thickness of about 400 μm to about 600 μm.

[0086] 14. The device of any one of embodiments 1-12, further comprising a third polymer layer having an average connectivity diameter of less than about 500 nm.

[0087] 15. The device of any one of embodiments 1-14, further comprising a port for introducing the plurality of cells into the lumen of the device.

[0088] 16. A device according to any one of embodiments 1-15, wherein the first polymer layer, the second polymer layer, the third polymer layer, or any combination thereof comprises a biocompatible polymer, and the biocompatible polymer is selected from: polycaprolactone (PCL), polyvinylidene fluoride (PVDF), polytetrafluoroethylene (PTFE), polyethylene (PE), methacrylate polymers, polyethyleneimine, polyethyleneimine-dextran sulfate, poly(vinylsiloxane)ecopolymere polyethyleneimine, phosphorylcholine, poly(ethyl methacrylate), polyurethane, poly(ethylene glycol) (PEG), poly(lactic-co-glycolic acid) (PLGA), hydroxyapatite, poly(lactic acid), polyhydroxyvalerate and copolymers thereof, polyhydroxybutyrate and copolymers thereof, polydioxanone, polyanhydrides, polycyanoacrylates, poly(amino acids), poly(orthoesters), polyesters, collagen, gelatin, cellulose polymers, chitosan, alginate, laminin, and any combination thereof.

[0089] 17. The device of embodiment 16, wherein the biocompatible polymer is polycaprolactone.

[0090] 18. A device according to any one of embodiments 1-17, wherein the plurality of cells are selected from: 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 from intestinal cells, pancreatic islets or pancreatic islet cells, Sertoli cells, β cells, pancreatic islet progenitor cells, β progenitor cells, peripheral blood progenitor cells, stem cells or their derivatives isolated from adult tissues, retinal progenitor cell-derived cells, cardiac progenitor cell-derived cells, bone progenitor cells, neuronal progenitor cells, genetically transformed cells, and any combination thereof.

[0091] 19. The device of any one of embodiments 1-18, wherein the first polymer layer and the second polymer layer are sealed directly along a periphery of the first polymer layer and the second polymer layer.

[0092] 20. The device of any one of embodiments 1-19, wherein the device does not comprise a support or frame.

[0093] 21. The device of any one of embodiments 1-20, wherein the first polymer layer, the second polymer layer, or both are non-laminated polymer layers.

[0094] 22. A method comprising: implanting a device according to any one of embodiments 1-21 into a subject suffering from or suspected of suffering from a disease or condition, wherein the device comprises a plurality of therapeutic cells, thereby treating the disease or condition.

[0095] 23. A method according to embodiment 22, wherein the subject is human.

[0096] 24. A method according to embodiment 22 or 23, wherein the plurality of cells are pancreatic islets or pancreatic islet cells.

[0097] 25. The method of any one of embodiments 22-24, wherein the subject has or is suspected of having diabetes.

[0098] 26. A method according to any one of embodiments 22-25, wherein the plurality of cells are allogeneic cells, xenogeneic cells or autologous cells.

[0099] 27. A method according to any one of embodiments 22-26, wherein the device is implanted together with a plurality of cells encapsulated within the lumen.

[0100] 28. A method according to any one of embodiments 22-27, wherein the device is implanted without any cells being encapsulated within the lumen.

[0101] 29. A method according to embodiment 28, wherein a plurality of cells are introduced into the lumen after the device is implanted in the subject for a period of time.

[0102] 30. The method of embodiment 29, wherein the period of time is at least 1 week.

[0103] 31. The method of any one of embodiments 22-30, wherein the therapeutic molecule is configured to be released from the device.

[0104] 32. The method of embodiment 31, wherein the therapeutic molecule diffuses out of the device.

[0105] 33. The method of embodiment 31 or 32, wherein the therapeutic molecule comprises insulin.

[0106] 34. The method of any one of embodiments 22-33, wherein the implantation is selected from the group consisting of: subcutaneous implantation, omental implantation, intraperitoneal implantation, retroperitoneal implantation, and intramuscular implantation.

[0107] 35. The method of any one of embodiments 22-34, wherein the device does not induce a foreign body response in the subject.

[0108] 36. The method of any one of embodiments 22-35, wherein the therapeutic cells are embedded within the second polymer layer.

[0109] 37. The method of any one of embodiments 22-36, wherein the therapeutic cells are attached to the second polymer layer.

[0110] Example

[0111] Example 1. Cell encapsulation device.

[0112] Figure 1A and Figure 1B Depicted are non-limiting examples of cell encapsulation devices according to embodiments of the present disclosure. Figure 1A A first polymer layer is depicted. The first polymer layer comprises a plurality of pores having an average connected diameter of about 200 nm and an average pore size of about 2 μm. Figure 1B The second polymer layer is depicted. The second polymer layer comprises a plurality of pores having an average pore diameter of about 500 μm to about 3 mm and an average connectivity diameter of about 100 μm to about 500 μm.

[0113] Example 2. Polymer Scaffold

[0114] The three-dimensional polymer scaffold was designed to facilitate islet transplantation in a manner that allows optimal packing of islets with better spatial distribution. The scaffold was designed with a random macroporous structure, which functions to allow islets to embed and blood vessels to grow within, so that the islets are vascularized ( Figure 2A-2E ).

[0115] Example 3. Cell encapsulation device.

[0116] The device is constructed with a nanoporous layer and a macroporous layer composed of polycaprolactone (e.g., the stent according to Example 2) ( Figure 3A-3C ). The nanoporous membrane is about 20 μm thick and consists of pores with a pore diameter of about 2 μm and a connectivity diameter of about 200 nm. The macroporous membrane is about 500 μm thick and consists of macroscopic pores with a connectivity diameter and pore diameter of 200-400 μm. To make the device, the macroporous membrane and the nanoporous membrane are stacked and sealed directly along the periphery.

[0117] Example 4. Implantation of syngeneic islets into diabetic mice

[0118] The device described in Example 3 was loaded with approximately 500 syngeneic C57BL / 6 islets, sealed, and implanted subcutaneously in C57BL / 6 mice ( Figure 5 ). On day 21, an intraperitoneal glucose tolerance test (IPGTT) was performed. The glucose tolerance test measures the clearance of an intraperitoneally injected glucose load from the body and can be used to detect glucose metabolism disorders that may be associated with human conditions such as diabetes or metabolic syndrome. The animals were fasted for approximately 16 hours, and fasting blood glucose levels were determined before a glucose solution was administered by intraperitoneal (IP) injection. Blood glucose levels were then measured at different time points during the subsequent 2 hours.

[0119] IPGTT was performed at 21 days with 500 islets encapsulated in the device implanted subcutaneously and 500 islets implanted directly into the renal capsule. The islets in the device had the same glucose profile as the islets in the renal capsule, indicating similar glucose responses ( Fig. 6A ).

[0120] In addition, insulin secretion was analyzed using a glucose-stimulated insulin secretion assay. The glucose stimulation index is a measure that quantifies beta cell function by comparing the ratio of insulin release in a high glucose state to that in a resting state. The ratio of insulin secreted under high to low glucose conditions is used to calculate the glucose stimulation index. Figure 6B As depicted in FIG. 5 , islets encapsulated within a device as described in Example 3 have a greater glucose stimulation index than islets implanted directly into the kidney capsule (KC).

[0121] Example 5. Device evaluation after implantation

[0122] The device was implanted subcutaneously for more than 30 days to allow colonization. After sacrifice, the tissue was excised and fixed at 4°C for more than 24 hours using Z-Fix and then washed in PBS. The tissue and device were imaged using CLARITY (Clear Lipid Exchange Acrylamide Hybridization Fine Imaging / Immunostaining / In Situ Hybridization Compatible Tissue Hydrogel), a technology that embeds tissue into a hydrogel matrix. Lipids are removed to produce an optically transparent mixture that is light-transmissive and molecularly permeable. The markers included are DAPI for nuclear staining, and CD31 and alpha smooth muscle actin (α-SMA) for staining blood vessels. Figure 7 Staining depicts blood vessels that have penetrated into and around the device.

[0123] Fig. 8A and Figure 8BDepicts immunohistochemical staining of sections after implantation of the device in mice. The image depicts pancreatic islets (arrows) embedded within the macroporous scaffold layer of the device.

[0124] Although preferred embodiments of the present invention have been shown and described herein, it is apparent to those skilled in the art that these embodiments are provided as examples only. Without departing from the present invention, those skilled in the art may conceive of many changes, variations and substitutions. It should be understood that the various alternatives of the embodiments of the present invention described herein may be used to implement the present invention. The following claims are intended to define the scope of the present invention, and are intended to cover methods and structures and their equivalents within the scope of these claims.

Claims

1. A device comprising: a) a first polymer layer comprising a plurality of pores having an average connected diameter of less than about 500 nm; b) a second polymer layer comprising a plurality of pores having an average connected diameter greater than about 100 μm; The first polymer layer and the second polymer layer define a lumen for encapsulating a plurality of cells. 2 . The device of claim 1 , wherein the plurality of pores present in the first polymer layer have an average connected diameter of about 10 nm to about 200 nm. 3 . The device of claim 1 , wherein the plurality of pores present in the first polymer layer have an average connected diameter of about 180 nm to about 220 nm. 4 . The device of claim 1 , wherein the plurality of pores present in the second polymer layer have an average connected diameter of about 100 μm to about 500 μm.

5. The device of any one of claims 1-3, wherein the plurality of pores present in the second polymer layer have an average connected diameter of about 200 μm to about 400 μm. 6 . The device of claim 1 , wherein the plurality of pores present in the first polymer layer have an average pore diameter of about 1 μm to about 5 μm.

7. The device of any one of claims 1-6, wherein the plurality of pores present in the second polymer layer have an average pore diameter of about 500 μm to about 3 mm.

8. The device of any one of claims 1-7, wherein the plurality of pores present in the second polymer layer have an average pore diameter of about 100 μm to about 1 mm.

9. The device of any one of claims 1-8, wherein the first polymer layer has a thickness of about 10 μm to about 200 μm.

10. A method comprising: Implanting a device according to any one of claims 1-9 into a subject suffering from or suspected of suffering from a disease or condition, wherein the device comprises a plurality of therapeutic cells, thereby treating the disease or condition.

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