Cell containing device

Abstract

The present disclosure provides a cell containment device having an array of channels to increase a surface area to volume ratio, and a method of manufacturing such a device.

Description

[0001] This application is a divisional application of the invention patent application of the same name 201880074042.6 filed on September 29, 2018.

[0002]

Cross reference

[0003] This application claims the benefit of U.S. Provisional Application No. 62 / 565,962, filed September 29, 2017, and U.S. Provisional Application No. 62 / 671,297, filed May 14, 2018, which are hereby incorporated herein by reference. [Background of the invention]

[0005] The therapeutic device that can deliver biological products is used to treat metabolic disorders, such as diabetes. The therapeutic device can be implantable to provide biological products, such as insulin, in the extended time period. These devices can include cell-containing devices and the matrix contained in the cell-containing device. The matrix can include cells for producing the biological products. As the size of the matrix increases, the availability of oxygen and other nutrients may be farther and less from the edge surface of the matrix, and there may be a region in the matrix where oxygen and nutrient concentrations are lower or there is no oxygen and nutrients. These regions where oxygen and nutrient concentrations are lower or there is no oxygen and nutrients may not support the cell viability and synthesis of biological products in the matrix. The spatial limitations in the delivery of oxygen, nutrients and other agents can limit the size of the device to the size that oxygen, nutrients and other agents can reach the cell. Therefore, it may be beneficial to improve the mass delivery to the internal region of the matrix of such devices and the matrix contained in the cell-containing device. [Summary of the invention]

[0006] The present disclosure relates generally to medical devices and methods. In various aspects, the present disclosure provides medical devices including cell containment devices, devices related thereto, and methods of making and using such devices.

[0007] In one aspect, described herein is a cell containment device comprising: a first membrane having a first surface comprising a plurality of channels and a plurality of second surfaces opposite the first surface; and a second membrane opposite the first membrane and attached to the plurality of second surfaces of the first membrane; wherein the first membrane and the second membrane form a closed compartment having a surface area to volume ratio of at least about 40 cm-1, and wherein the closed compartment provides a volume for containing cells within the device.

[0008] In some embodiments, the compartment comprises a single continuous open space. In some embodiments, the volume is about 8 μl to about 1,000 μl. In some embodiments, at least one of the length and width of the device is about 0.25 cm to about 3 cm. In some embodiments, the thickness of the device is at least about 300 μm. In some embodiments, the plurality of channels are substantially perpendicular to the first membrane. In some embodiments, the plurality of channels are arranged in a linear array. In some embodiments, the plurality of channels are arranged in an annular array. In some embodiments, the average diameter of the channel is about 400 μm to about 3,000 μm. In some embodiments, the diameter is measured at the narrowest point in the channel. In some embodiments, the center of each channel is separated from the center of another channel by a distance of about 75 μm to about 500 μm. In some embodiments, the height to diameter ratio of the channel is at least about 0.2. In some embodiments, the number of channels per unit area of ​​the device along the transverse plane is greater than about 50 / cm2. In some embodiments, at least one of the first membrane and the second membrane includes a plurality of nodes interconnected by a plurality of fibrils. In some embodiments, at least one of the first film and the second film includes PVDF, PTFE, ePTFE, PCL, PE / PES, PP, PS, PMMA, PLGA, PLLA or any combination thereof. In some embodiments, the device also includes an opening that runs through the first film and the second film in the channel. In some embodiments, the concentricity of the opening relative to the channel is at most 25% of the diameter of the channel. In some embodiments, the device also includes a frame configured to receive the device. In some embodiments, the framework is configured to receive multiple cell accommodating devices. In some embodiments, the framework includes a flexure mechanism, and the flexure mechanism is configured to prevent the buckling of the cell accommodating device. In some embodiments, the device also includes a cell colony. In some embodiments, the cell colony is an insulin secretion colony. In some embodiments, the cell colony is a stem cell-derived cell capable of achieving glucose-stimulated insulin secretion (GSIS). In some embodiments, the device also includes a coating, and the coating includes a hydrophilic polymer. In some embodiments, the insulin diffusion coefficient of the device is about 2x10^-6cm2 / s to about 1x10^-5cm2 / s. In some embodiments, the maximum insulin diffusion distance of the device is less than about 150μm. In some embodiments, the first membrane and the second membrane are fused by a fusion peeling force of at least about 0.4N. In some embodiments, at least one of the first membrane and the second membrane is semi-permeable. In some embodiments, the semi-permeability of the first membrane, the second membrane, or both is configured to protect the cells from immune attack.In some embodiments, the semipermeability of the first membrane, the second membrane, or both is configured to protect the cells from immune attack in the absence of immunosuppressive therapy. In some embodiments, at least one of the first membrane and the second membrane is configured to achieve vascularization of the cells within the device. In some embodiments, at least one of the first membrane and the second membrane is configured to achieve vascularization of the cells within the device in the absence of immunosuppressive therapy.

[0009] Another aspect provided herein is a cell-containing device comprising: a first membrane having a first surface comprising a plurality of channels and a plurality of second surfaces opposite the first surface; and a second membrane opposite the first membrane and attached to the plurality of second surfaces of the first membrane; wherein the first membrane and the second membrane form a closed compartment, wherein the closed compartment provides a volume for containing 1 million to 1 billion insulin-producing cells within the device, and wherein the membrane allows insulin to diffuse from the device while retaining the insulin-producing cells within the device.

[0010] Another aspect provided herein is a composition comprising insulin-producing cells and a device for accommodating the insulin-producing cells, wherein the device releases insulin while the insulin-producing cells are maintained in the device after being implanted in an individual body, and promotes tissue vascularization in and around the device. In some embodiments, an immunosuppressant is not administered to an individual during implantation or vascularization of the device. In some embodiments, the device comprises 1,000,000 to 1,000,000,000 insulin-producing cells. In some embodiments, the thickness of the device is at least about 300 μm. In some embodiments, the device comprises a film comprising a plurality of nodes interconnected by a plurality of fibrils.

[0011] In another aspect, a method for manufacturing a cell-containing device is described herein, comprising: providing a first membrane having a first side and an opposing second side; forming a plurality of channels within the first side of the first membrane; and fusing a second membrane to the second side of the first membrane to form a compartment for containing cells between the second side of the first membrane and the second membrane.

[0012] In some embodiments, forming a plurality of channels in the first film comprises: heating the first film for a predetermined time at a predetermined pressure and a predetermined temperature; and molding the plurality of channels with a mold. In some embodiments, the fusion of the second film to the first film is performed in the mold. In some embodiments, the mold comprises a male mold. In some embodiments, the mold comprises a female mold. In some embodiments, the predetermined temperature is about 100°C to about 600°C. In some embodiments, the predetermined pressure is about 2 pounds per square inch (psi) to about 140psi. In some embodiments, the predetermined time is about 3 minutes to about 30 minutes. In some embodiments, the predetermined pressure is about 3.5psi, and wherein the predetermined temperature is about 370°C. In some embodiments, forming a plurality of channels in the first film and fusing the second film to the first film comprises: placing the first film and the second film in a frame, wherein the first film and the second film are substantially parallel, substantially aligned, and separated by a gap distance; and striking one or more points on the first film with a fusion tool, wherein the fusion tool is heated to a set fusion temperature, and wherein the fusion tool is in contact with the film for a set fusion time during each strike. In some embodiments, striking the first film pierces the first film, the second film, or both, and fuses a portion of the first film to the second film. In some embodiments, the frame surrounds at least a portion of the outer edges of the first film and the second film. In some embodiments, the gap distance is about 300 μm to about 1,200 μm. In some embodiments, the impact contact area of ​​the fusion tool is at least about 0.07 mm2. In some embodiments, the impact of the one or more points on the first film with the fusion tool includes striking each of the one or more points up to about 16 times. In some embodiments, the impact of the one or more points on the first film with the fusion tool includes striking each of the one or more points 1 to 6 times. In some embodiments, the set fusion temperature is about 250°C to about 600°C. In some embodiments, the set fusion time is less than about 1 second. In some embodiments, at least one of the first film and the second film is substantially flat. In some embodiments, the method further comprises embossing the first film before forming the plurality of channels in the first film. In some embodiments, the method further comprises laser ablation of a portion of the first film and the second film in the plurality of channels. In some embodiments, the laser ablation removes the fused portion of the first film and the second film to form an opening. In some embodiments, the concentricity of the opening relative to the channel is at most 25% of the diameter of the channel.In some embodiments, at least one of the first film and the second film comprises PVDF, PTFE, ePTFE, PCL, PE / PES, PP, PS, PMMA, PLGA, PLLA or any combination thereof. In some embodiments, the method further comprises coating the device with a hydrophilic polymer. In some embodiments, the first film is sintered. In some embodiments, the second film is not sintered. In some embodiments, the second film and the first film are fused with a fusion peeling force of at least about 0.2N.

[0013] Another aspect provided herein is a method comprising: contacting tissue of a diabetic or prediabetic subject with a device comprising a population of insulin-secreting cells, wherein the device comprises: a first membrane having a first surface comprising a plurality of channels and a plurality of second surfaces opposite the first surface; and a second membrane opposite the first membrane and attached to the plurality of second surfaces of the first membrane; wherein the first membrane and the second membrane form a closed compartment having a surface area to volume ratio of at least about 40 cm-1, and wherein the closed compartment provides a volume for accommodating cells within the device; and releasing insulin from the population of insulin-secreting cells in response to an elevated blood glucose level in the diabetic subject, wherein the elevated blood glucose level is higher than the blood glucose level in a non-diabetic subject.

[0014] In some embodiments, the insulin-secreting cell colony releases insulin in an amount sufficient to reduce the blood glucose level in the diabetic or prediabetic subject. In some embodiments, the release of insulin stops when the blood glucose level in the diabetic subject is reduced to a normal level. In some embodiments, the release of insulin restarts when the insulin-secreting cell colony is exposed to the elevated blood glucose level in the diabetic subject again. In some embodiments, the insulin-secreting cell colony is a stem cell-derived cell colony. In some embodiments, the insulin-secreting cell colony is capable of achieving glucose-stimulated insulin secretion (GSIS). In some embodiments, at least one of the first and second membranes is semipermeable. In some embodiments, the semipermeability of the first membrane, the second membrane, or both is configured to protect the cells from immune attack. In some embodiments, the semipermeability of the first membrane, the second membrane, or both is configured to protect the cells from immune attack in the absence of immunosuppressive therapy. In some embodiments, at least one of the first and second membranes is configured to achieve vascularization of the cells in the device. In some embodiments, at least one of the first and second membranes is configured to achieve vascularization of the cells in the device in the absence of immunosuppressive therapy.

[0015] In one aspect, a cell containment device is described herein, comprising: a first surface, the first surface defining an outer surface of the device and having a surface area; a second surface, the second surface opposite the first surface, wherein the second surface defines an inner surface of the device; and a compartment, the compartment being enclosed within the second surface, wherein the compartment provides a volume for containing cells within the device; wherein the ratio of the surface area to the volume is equal to or greater than 50 cm -1 . In some aspects, the device comprises a plurality of channels extending through a transverse plane of the device. In some aspects, each of the plurality of channels comprises a diameter equal to or greater than 400 μm. In some aspects, the diameter is measured at the narrowest point in the channel. In some aspects, each of the plurality of channels is separated from each other by a distance of no more than 450 μm. In some aspects, each of the plurality of channels comprises a height to diameter ratio equal to or greater than 0.2. In some aspects, the number of channels per unit area measured along the transverse plane of the device is greater than 50 / cm 2 In some aspects, the number of channels per unit area measured along a transverse plane of the device is greater than 100 / cm 2 In some aspects, the ratio of the surface area to the volume is greater than 80 cm -1In some aspects, the ratio of the surface area to the volume is greater than 100 cm -1 In some aspects, the ratio of the surface area to the volume is greater than 120 cm -1 . In some aspects, the device includes a single continuous open space having the volume. In some aspects, the first surface or the second surface includes a plurality of nodes interconnected by a plurality of fibrils. In some aspects, the device includes a thickness greater than 300 μm measured along the transverse plane of the device. In some aspects, the first surface or the second surface includes PVDF, PTFE, ePTFE, PCL, PE / PES, PP, PS, PMMA, PLGA or PLLA. In some aspects, the device also includes a frame, wherein the frame is configured to accommodate the device. In some aspects, the frame is configured to accommodate a plurality of cell-containing devices. In some aspects, the frame includes a flexure mechanism to prevent the buckling of the cell-containing device. In some aspects, the device also includes a cell colony. In some aspects, the cell colony is an insulin secreting colony. In some aspects, the cell colony is a stem cell-derived cell capable of achieving glucose-stimulated insulin secretion (GSIS).

[0016] In some aspects, the device further comprises a coating having a hydrophilic polymer. In some aspects, the volume for accommodating the cells is inversely proportional to at least one of the diameter of the plurality of channels and the number of channels per unit area of ​​the device. In some aspects, the ratio of the surface area of ​​the device to the volume is proportional to at least one of the diameter of the plurality of channels and the number of channels per unit area of ​​the device. In some aspects, the ratio of the surface area of ​​the device to the volume achieves a greater mass transport into and / or out of the device. In other aspects, a cell-containing device is described herein, comprising: a substrate; a top surface opposite to the substrate; a height, the height extending from the substrate to the top surface along the transverse plane of the device, wherein the height is greater than 300 μm; a compartment for accommodating cells, wherein the compartment is enclosed between the substrate and the top surface; and a plurality of channels, the plurality of channels extending along the transverse plane of the device, wherein the maximum oxygen diffusion distance of the device is less than 150 μm. In some aspects, the device comprises a height greater than 600 μm. In some aspects, the substrate is substantially flat. In some aspects, each of the plurality of channels comprises a diameter equal to or greater than 400 μm. In some aspects, each of the plurality of channels is separated from each other by a distance of no more than 450 μm. In some aspects, each of the plurality of channels comprises a diameter equal to or greater than 400 μm. In some aspects, the diameter is measured at the narrowest point in the channel. In some aspects, each of the plurality of channels is separated from each other by a distance of no more than 450 μm. In some aspects, each of the plurality of channels comprises a height to diameter ratio equal to or greater than 0.2. In some aspects, the number of channels per unit area measured along a transverse plane of the device is greater than 50 / cm 2 In some aspects, the number of channels per unit area measured along a transverse plane of the device is greater than 100 / cm 2. In some aspects, the device includes a single continuous compartment for accommodating cells. In some aspects, the first surface or the second surface includes a plurality of nodes interconnected by a plurality of fibrils. In some aspects, the first surface or the second surface includes PVDF, PTFE, ePTFE, PCL, PE / PES, PP, PS, PMMA, PLGA or PLLA. In some aspects, the device also includes a frame, wherein the frame is configured to accommodate the device. In some aspects, the frame is configured to accommodate multiple cell accommodating devices. In some aspects, the frame includes a flexure mechanism to prevent the buckling of the cell accommodating device. In some aspects, the device also includes a cell colony. In some aspects, the cell colony is an insulin secreting colony. In some aspects, the cell colony is a stem cell-derived cell capable of achieving glucose-stimulated insulin secretion (GSIS). In some aspects, the device also includes a coating with a hydrophilic polymer. In some aspects, the volume for accommodating the cells is inversely proportional to at least one of the diameter of the multiple channels and the number of channels per unit area of ​​the device. In some aspects, the ratio of the surface area to the volume of the device is proportional to at least one of the diameter of the plurality of channels and the number of channels per unit area of ​​the device. In some aspects, the ratio of the surface area to the volume of the device is proportional to at least one of the diameter of the plurality of channels and the number of channels per unit area of ​​the device. In some aspects, a larger ratio of the surface area to the volume of the device enables greater mass transport into and / or out of the device.

[0017] In other aspects, a method for manufacturing a cell-containing device is described herein, the method comprising: providing a first membrane; adjusting the temperature and / or pressure around the first membrane to a predetermined value; deforming the first membrane; and fusing a second membrane to the first membrane, wherein the compartment between the first membrane and the second membrane defines a compartment for containing cells. In some aspects, the predetermined value is less than 170°C. In some aspects, the predetermined value is less than 140 pounds per square inch (psi). In some aspects, the predetermined value is less than 370°C. In some aspects, the predetermined value is less than 5psi. In some aspects, deforming the first membrane includes depressing a portion of the first membrane with a tool. In some aspects, the tool comprises: a substantially flat surface, the surface being configured to be parallel to the first membrane; and a plurality of protrusions on the surface, the protrusions being configured to depress a portion of the first membrane. In some aspects, each of the plurality of protrusions comprises a cylinder. In some aspects, the tool comprises a tip, wherein the tip has a contact area at the free end. In some aspects, the contact area is equal to or greater than 0.07mm 2. In some aspects, the fusion is performed with the tip, wherein the tip presses the first film and the second film into contact with each other for a predetermined time. In some aspects, the deformation and fusion are performed in one step with the tip, wherein the tip contacts the first film, moves vertically away from the first film toward the second film, and presses the first film and the second film into contact with each other for a predetermined time. In some aspects, the tip is adjusted to a predetermined temperature value. In some aspects, the tip is pressed with a predetermined pressure value. In some aspects, the predetermined time is equal to or greater than 1 second. In some aspects, the cylinder includes a diameter equal to or greater than 300 μm. In some aspects, the cylinder includes a height equal to or greater than 300 μm. In some aspects, deforming the first film is completed without causing rupture of the film. In some aspects, deforming the first film includes forming a plurality of features on the film. In some aspects, each of the plurality of features includes a diameter equal to or greater than 300 μm. In some aspects, each of the plurality of features includes a depth equal to or greater than 300 μm. In some aspects, the method further comprises adjusting the temperature and / or pressure around the first film to control the characteristics of the feature. In some aspects, increasing the temperature and / or pressure around the first film increases the depth of the feature. In some aspects, fusing the second film to the first film comprises fusing the second film and the first film into a continuous layer. In some aspects, fusing the second film to the first film is implemented at a temperature and / or pressure having a second predetermined value. In some aspects, the second predetermined value is less than 230° C. In some aspects, the second film is substantially flat. In some aspects, after deformation, the first film is embossed. In some aspects, after fusion, the device has a substantially flat surface and a convex surface opposite to the substantially flat surface. In some aspects, the method further comprises removing the fused portion of the first film and the second film via laser ablation, thereby forming a channel across the device. In some aspects, the method further comprises mounting the device on a frame. In some aspects, the method further comprises implanting the device on the frame into a subject. In some aspects, the method further comprises encapsulating the cells in the compartment. In some aspects, the method further comprises implanting the device into a subject. In some aspects, the first film or the second film comprises PVDF, PTFE, ePTFE, PCL, PE / PES, PP, PS, PMMA, PLGA or PLLA. In some aspects, the method further comprises coating the device with a hydrophilic polymer. In some aspects, the first film is sintered. In some aspects, the second film is not sintered.

[0018] In other aspects, a cell-containing device is described herein, comprising: a substrate; a top surface opposite to the substrate; a height extending from the substrate to the top surface along the transverse plane of the device, wherein the height is equal to or less than 300 μm; and a compartment for accommodating cells, wherein the compartment is enclosed between the substrate and the top surface. In some aspects, the height is less than 250 μm. In some aspects, the substrate comprises a sintered film. In some aspects, the top surface comprises a sintered film. In some aspects, the substrate comprises a coating film, wherein the coating increases the hydrophilicity of the film. In some aspects, the device comprises at least one fusion point, wherein the point comprises a portion of the substrate fused to a portion of the top surface corresponding to the portion of the substrate and wherein the point is configured to limit the change in the height. In some aspects, the diameter of the point is about 0.5 mm to about 3 mm. In some aspects, the diameter of the point is at least about 0.5 mm. In some aspects, the diameter of the point is at most about 3 mm.In some aspects, the diameter of the dots is about 0.5 mm to about 0.75 mm, about 0.5 mm to about 1 mm, about 0.5 mm to about 1.25 mm, about 0.5 mm to about 1.5 mm, about 0.5 mm to about 1.75 mm, about 0.5 mm to about 2 mm, about 0.5 mm to about 2.25 mm, about 0.5 mm to about 2.5 mm, about 0.5 mm to about 2.75 mm, about 0.5 mm to about 3 mm, about 0.75 mm to about 1 mm, about 0.75 mm to about 1.25 mm, about 0.75 mm to about 1. 5mm, about 0.75mm to about 1.75mm, about 0.75mm to about 2mm, about 0.75mm to about 2.25mm, about 0.75mm to about 2.5mm, about 0.75mm to about 2.75mm, about 0.75mm to about 3mm, about 1mm to about 1.25mm, about 1mm to about 1.5mm, about 1mm to about 1.75mm, about 1mm to about 2mm, about 1mm to about 2.25mm, about 1mm to about 2.5mm, about 1mm to about 2.75mm, about 1mm to about 3mm, about 1.25 mm to about 1.5 mm, about 1.25 mm to about 1.75 mm, about 1.25 mm to about 2 mm, about 1.25 mm to about 2.25 mm, about 1.25 mm to about 2.5 mm, about 1.25 mm to about 2.75 mm, about 1.25 mm to about 3 mm, about 1.5 mm to about 1.75 mm, about 1.5 mm to about 2 mm, about 1.5 mm to about 2.25 mm, about 1.5 mm to about 2.5 mm, about 1.5 mm to about 2.75 mm, about 1.25 mm to about 3 mm, about 1.5 mm to about 1.75 mm, about 1.5 mm to about 2 mm, about 1.5 mm to about 2.5 mm, about 1.5 mm to about 2.75 mm, about 1.5 mm to about 3 mm, about 1.75 mm to about 2 mm In some aspects, the diameter of the point is about 0.5 mm, about 0.75 mm, about 1 mm, about 1.25 mm, about 1.5 mm, about 1.75 mm, about 2 mm, about 2.25 mm, about 2 mm to about 2.5 mm, about 2 mm to about 2.75 mm, about 2 mm to about 3 mm, about 2.25 mm to about 2.5 mm, about 2.25 mm to about 2.75 mm, about 2.25 mm to about 3 mm, about 2.5 mm to about 2.75 mm, about 2.5 mm to about 3 mm, or about 2.75 mm to about 3 mm. In some aspects, the diameter of the point is about 0.5 mm, about 0.75 mm, about 1 mm, about 1.25 mm, about 1.5 mm, about 1.75 mm, about 2 mm, about 2.25 mm, about 2.5 mm, about 2.75 mm, or about 3 mm. In some aspects, the point is at least 3 mm away from another fusion point. In some aspects, the dots are formed with an adhesive placed between the portion of the substrate and the portion of the top surface. In some aspects, the volume of the compartment is inversely proportional to at least one of a diameter of the dots and a number of dots per unit area of ​​the device.In some aspects, the ratio of the surface area of ​​the device to the volume of the device is proportional to at least one of the diameter of the dots and the number of dots per unit area of ​​the device. In some aspects, a larger ratio of the surface area to volume of the device enables greater mass transport into and / or out of the device.

[0019] In other aspects, a method is described herein, comprising: a) contacting a tissue of a diabetic subject with a device comprising a population of insulin-secreting cells, wherein the device comprises: a first surface, the first surface defining an exterior surface of the device and having a surface area; a second surface, the second surface opposite the first surface, wherein the second surface defines an interior surface of the device; and a compartment enclosed within the second surface, wherein the compartment provides a volume for containing cells within the device; wherein the ratio of the surface area to the volume is equal to or greater than 50 cm -1 ; And b) release insulin from the insulin secreting cell colony in response to the blood glucose level of the increase in the diabetic subject, wherein the blood glucose level of the increase is higher than the blood glucose level in the non-diabetic subject. In some aspects, the insulin secreting cell colony releases the insulin of the amount sufficient to reduce the blood glucose level in the diabetic subject. In some aspects, the release of insulin stops when the blood glucose level in the diabetic subject is reduced to normal levels. In some aspects, the release of insulin restarts when the insulin secreting cell colony is exposed to the blood glucose level of the increase in the diabetic subject again. In some aspects, the insulin secreting cell colony is a stem cell derived cell colony. In some aspects, the insulin secreting cell colony can achieve glucose stimulated insulin secretion (GSIS).

[0020] In other aspects, a method is described herein, comprising: a) contacting a diabetic subject's tissue with a device comprising a population of insulin-secreting cells, wherein the device comprises: a substrate; a top surface opposite the substrate; a height extending from the substrate to the top surface along a transverse plane of the device, wherein the height is greater than 300 μm; a compartment for accommodating cells, wherein the compartment is enclosed between the substrate and the top surface; and a plurality of channels extending along the transverse plane of the device, wherein the maximum oxygen diffusion distance of the device is less than 150 μm; and b) releasing insulin from the insulin-secreting cell population in response to an elevated blood glucose level in the diabetic subject, wherein the elevated blood glucose level is higher than the blood glucose level in a non-diabetic subject. In some aspects, the insulin-secreting cell population releases insulin in an amount sufficient to reduce the blood glucose level in the diabetic subject. In some aspects, the release of insulin stops when the blood glucose level in the diabetic subject is reduced to a normal level. In some aspects, the release of insulin resumes when the insulin-secreting cell population is exposed to an elevated blood glucose level in the diabetic subject again. In some aspects, the insulin secreting cell colony is a stem cell derived cell colony.In some aspects, the insulin secreting cell colony is capable of achieving glucose stimulated insulin secretion (GSIS).

[0021] In other aspects, a method is described herein, comprising: a) contacting a diabetic subject's tissue with a device comprising an insulin-secreting cell population, wherein the device comprises: a substrate; a top surface opposite the substrate; a height extending from the substrate to the top surface along a transverse plane of the device, wherein the height is equal to or less than 300 μm; and a compartment for accommodating cells, wherein the compartment is enclosed between the substrate and the top surface; and b) releasing insulin from the insulin-secreting cell population in response to an elevated blood glucose level in the diabetic subject, wherein the elevated blood glucose level is higher than the blood glucose level in a non-diabetic subject. In some aspects, the insulin-secreting cell population releases insulin in an amount sufficient to reduce the blood glucose level in the diabetic subject. In some aspects, the release of insulin stops when the blood glucose level in the diabetic subject is reduced to a normal level. In some aspects, the release of insulin resumes when the insulin-secreting cell population is exposed to an elevated blood glucose level in the diabetic subject again. In some aspects, the insulin-secreting cell population is a stem cell-derived cell population. In some aspects, the population of insulin-secreting cells is capable of glucose-stimulated insulin secretion (GSIS).

[0022] [Incorporated by reference]

[0023] 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

[0024] The novel features of the present disclosure are particularly set forth in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description, which sets forth illustrative embodiments in which the principles of the present disclosure are utilized, and the accompanying drawings of the present disclosure:

[0025] Figure 1 High magnification images showing polyvinylidene fluoride (PVDF) cell containment devices according to some embodiments;

[0026] Figure 2 It is demonstrated according to some embodiments that the amount of cells loaded into the device varies with the spot spacing and the presence of constraints.

[0027] Figure 3 Showing regions with different oxygen pressures within a bi-layer flat sheet and channel array device according to some embodiments;

[0028] Figure 4 The surface area and vascularization potential of a device having a flat configuration and a device having an array of channels are shown according to some embodiments;

[0029] Figure 5 Devices having a flat configuration or devices having an array of channels and increased vascularization are shown according to some embodiments;

[0030] Figure 6 Computer-aided design (CAD) rendering of a device showing arrays of hexagonal channels of varying sizes, according to some embodiments.

[0031] Figure 7 A CAD rendering of a cell containment device and a scanning electron micrograph of a cross-section of the cell containment device are shown according to some embodiments;

[0032] Figure 8 Deformation of the membrane from a flat configuration to a shaped configuration having channels is shown according to some embodiments;

[0033] Fig. 9 According to some embodiments, fusion of the first film with the flat second film after the deforming step and images of the fused first and second films are shown.

[0034] Fig.10Scanning electron micrographs and higher magnification scanning electron micrographs showing cross-sections of a cell containment device along various portions of the cell containment device according to some embodiments;

[0035] Fig.11 Scanning electron micrographs showing low and high magnification of a cross-section of a cell containment device at the interface of a fused first membrane and a second membrane according to some embodiments;

[0036] Fig.12 Scanning electron micrographs showing cross-sections of polyvinylidene fluoride (PVDF) membranes after undergoing a deformation step at various temperatures and pressures according to some embodiments;

[0037] Fig.13 Scanning electron micrographs showing cross-sections of a cell containment device after undergoing a fusion step at various temperatures and pressures according to some embodiments;

[0038] Fig.14 Presentation of cell containment devices having scalloped perimeters and variations in channel dimensions to achieve various SA:V ratios is shown according to some embodiments.

[0039] Fig.15 Heat flow measurements of PVDF membranes are shown according to some embodiments before processing, after processing at 173°C and 100 psi with rapid cooling, and after processing at 160°C and 100 psi;

[0040] Fig.16 Scanning electron micrographs of cross-sections of cell containment devices are shown according to some embodiments, wherein the deforming step is performed under different pressure and temperature conditions.

[0041] Fig.17 According to some embodiments measured channel depths in a cell containment device are shown, wherein the deforming step is performed under different pressure and temperature conditions.

[0042] Fig.18 Surface interferometry profiling of a cell containment device is presented according to some embodiments.

[0043] Fig.19 showing low and high magnification scanning electron micrographs of a first film of expanded polytetrafluoroethylene (ePTFE) after a deformation step performed at various temperatures and pressures according to some embodiments;

[0044] Fig. 20 Scanning electron micrographs of ePTFE cell containment devices after a deformation step performed at 360°C and 6 psi and a fusion step performed at 370°C for 5 minutes are shown according to some embodiments;

[0045] Fig.21 Scanning electron micrographs and images showing a cell containment device having a lumen of a fused portion cut by laser ablation according to some embodiments;

[0046] Fig. 22 A scanning electron micrograph showing a cross-section of a cell containment device after fabrication according to some embodiments;

[0047] Fig.23 According to some embodiments, a design of a macro device equipped with a hexagonal channel array device and equipped with a flexure mechanism is shown;

[0048] Fig.24 Filling of a cell-containing device with a matrix containing cells is shown in a configuration where the device is mounted to a mechanical frame for support (left) and in a configuration using an external filling tube and a frameless design (right), according to some embodiments.

[0049] Fig.25 According to some embodiments a hematoxylin and eosin (H&E) stained tissue section is shown of a cell containment device filled with a matrix having cells.

[0050] Fig.26 Demonstrating vasculature surrounding a cell containment device 20 days after implantation in a rat according to some embodiments;

[0051] Fig. 27 Demonstrating vasculature surrounding a cell containment device 90 days after implantation in a rat according to some embodiments;

[0052] Fig.28 According to some embodiments, preperitoneal, intraomental, suprahepatic, and subcutaneous implantation of cell containment devices in rats is demonstrated;

[0053] Fig.29 H&E stained tissue sections showing vascularization around the cell containment device and vasculature in the channels after in vivo implantation in rats according to some embodiments;

[0054] Fig.30 Demonstrating vascularization around cell containment devices having low channel density and high channel density after in vivo implantation according to some embodiments;

[0055] Fig.31 Demonstrating the number of blood vessels and blood vessel branches per device observed in cell-containing devices having low channel density and high channel density after in vivo implantation according to some embodiments;

[0056] Fig.32The effect of device design and surface area to volume ratio on insulin response is shown according to some embodiments;

[0057] Fig.33 The surface area to volume ratio achievable by variation of channel density and the estimated permissible cluster diameter for channels with a diameter of 500 μm are shown according to some embodiments;

[0058] Fig.34 A schematic diagram showing a fusion process by spot welding two films using a tip according to some embodiments;

[0059] Fig.35 Demonstrating the breaking load of two flat ePTFE membranes fused at different temperatures and for different lengths of time according to some embodiments;

[0060] Fig.36 Scanning electron micrograph showing the edge of a device sealed in 0.5 seconds at 345°C according to some embodiments;

[0061] Fig.37 Stress-strain curves showing exemplary peel tests according to some embodiments;

[0062] Fig.38 An ePTFE cell containment device after hydrophilic coating treatment in water is shown according to some embodiments;

[0063] Fig.39 A graph showing the failure fill pressure (psi) of an ePTFE cell containment device without a frame that was filled until bursting, and a PVDF and ePTFE cell containment device prototype, according to some embodiments;

[0064] Fig.40 Prototypes showing three cell containment devices assembled onto a frame for human use according to some embodiments;

[0065] Fig.41 An example of a protocol for forming a hydrophilic coating on the surface of a membrane is presented according to some embodiments;

[0066] Fig.42 showing H&E stained tissue sections where the continuous interior spaces of ePTFE and PVDF cell containment devices are completely filled with cells, according to some embodiments;

[0067] Fig.43 An image showing an ePTFE cell containment device filled to maximum capacity with cells according to some embodiments;

[0068] Fig.44showing H&E stained tissue sections of a PVDF cell containment device with cells 90 days after in vivo implantation at the preperitoneal site in a rat according to some embodiments;

[0069] Fig.45 Showing H&E stained tissue sections of PVDF cell containment devices populated with SC islet cells 90 days after implantation in subcutaneous and preperitoneal sites in rats according to some embodiments;

[0070] Fig.46 An example of a macro device including three ultrathin devices is shown according to some embodiments;

[0071] Fig.47 Scanning electron micrographs showing different ePTFE membranes that can be used in ultrathin devices according to some embodiments;

[0072] Fig.48 Demonstrating, according to some embodiments, the flux of C-peptide in an implanted high-throughput ultrathin device having a selectively permeable membrane and populated with rat islet cells in response to 20 mM glucose stimulation;

[0073] Fig.49 According to some embodiments, static C-peptide release in an ultrathin device with a high-flux membrane or a selectively permeable membrane and filled with encapsulated islet cells is demonstrated before (LG or low glucose), during (HG), and after (LG) high glucose stimulation;

[0074] Fig.50 Demonstrating dynamic GSIS (glucose stimulated insulin secretion) of ultrathin devices with AS-1 membranes according to some embodiments;

[0075] Fig.51 Scanning electron micrographs showing non-sintered and sintered films according to some embodiments and changes in the microstructure of the films, wherein there is fusion and coalescence of nodes and fibrils of the films;

[0076] Fig.52 According to some embodiments, the results are close to 1x10 -6 mol / m 2 Comparison of hydrophilic coating methods for membranes with a target insulin flux of / s;

[0077] Fig.53 Showing membranes coated by three different coating methods V1, V2 and V3 according to some embodiments, the membranes have been stained with H&E dye as an indication of the hydrophilicity of the membrane;

[0078] Fig.54 Demonstrating hydraulic permeability of a membrane, a membrane subjected to a V1 coating process, and a membrane subjected to a V3 coating process according to some embodiments;

[0079] Fig.55 A setup for scaling up a coating method and drying and wetting a coated film is presented according to some embodiments;

[0080] Fig.56 Simulations of the mechanical properties of a 300 μm thick PEEK frame for two cell containment devices under a force of 150 mN are shown according to some embodiments;

[0081] Fig.57 An example of a PEEK frame holding a cell containment device having a fusion point at the center of the device is shown according to some embodiments;

[0082] Fig.58 showing an image of a single frame module having a single cell receptacle on the frame with a centered fusion point, wherein the single cell receptacle is maximally filled, according to some embodiments;

[0083] Fig.59 According to some embodiments, H&E stained tissue sections are shown when stored in a maximally filled cell containment device under standard conditions of 20% oxygen and 37°C for 2 days, 4 days, 8 days, and 10 days, wherein high cell viability is maintained after 10 days of storage;

[0084] Fig.60 According to some embodiments, blood glucose levels were demonstrated over 90 days in an ultrathin cell containment device with AS-1 membrane and filled with SC-islet cells implanted in a NOD scid gamma (NSG) mouse model (immunodeficient mouse model);

[0085] Fig.61A An image showing an ultrathin device having a high-throughput ePTFE membrane with a hydrophilic coating and filled with 8 million SC islet cells according to some embodiments;

[0086] Fig.61B H&E staining images showing high cell density within the device (including the core of the device) 90 days after implantation in the NSG mouse model according to some embodiments;

[0087] Fig.61C H&E staining images showing high cell density within the device (including the core of the device) 90 days after implantation in the NSG mouse model according to some embodiments;

[0088] Fig.62showing low magnification images of tissue sections of cell-filled ultrathin devices having AS-1 membranes and filled with SEM-01 cells 30 days after implantation in a mouse model in vivo according to some embodiments;

[0089] Fig.63 High magnification images of tissue sections showing cell-filled ultrathin devices having AS-1 membranes and filled with SEM-01 cells 30 days after implantation in a mouse model in vivo according to some embodiments;

[0090] Fig.64 According to some embodiments, microaggregates of endocrine cells are shown prior to encapsulation and filling into a cell containment device;

[0091] Fig.65 Showing stained tissue images of ultrathin devices populated with microclusters of endocrine cells 3 months after implantation in mice according to some embodiments;

[0092] Fig.66 According to some embodiments, serum C-peptide levels and total insulin content of an ultrathin device implanted with endocrine cells or filled with endocrine cells are displayed;

[0093] Fig.67 Showing H&E stained images of explants of ultrathin devices comprising a coated selectively permeable membrane and filled with SC-islet cells after 3 months at a preperitoneal site in nude mice according to some embodiments;

[0094] Fig.68 Showing H&E staining images of an ultrathin device populated with 16 million SC-islet cells 12 weeks after implantation at the preperitoneal site in a nude mouse model according to some embodiments;

[0095] Fig.69 showing serum C-peptide levels over 60 minutes in 4 different mice implanted with ultrathin devices populated with SC-islet cells according to some embodiments;

[0096] Fig.70A and Fig.70B showing H&E staining images of empty ultrathin devices with AS-1 membrane implanted in an immunocompetent Black 6 mouse model, which do not induce a foreign body response (FBR), according to some embodiments;

[0097] Fig.71 H&E stained images showing ultrathin devices with viable, intact rat islet cells after 90 days in vivo according to some embodiments;

[0098] Fig.72According to some embodiments, blood glucose levels before, during, and 90 days after implantation of the ultrathin device in a diabetic mouse model are shown;

[0099] Fig.73 According to some embodiments, an ultrathin device designed for implantation in a mouse is presented, wherein the device has a fusion point at the center;

[0100] Fig.74A and Fig.74B Various configurations of a macro device frame for holding a plurality of cell-receiving devices are shown according to some embodiments;

[0101] Fig.75A Ultrathin devices for human implantation having arrays of fusion sites that provide adhesive constraints are presented according to some embodiments;

[0102] Fig.75B According to some embodiments, a setup with a porous metal press plate to provide external constraints for the ultra-thin device is shown;

[0103] Fig.76A Demonstrating the configuration of an ultrathin device with adhesive constraints.

[0104] Fig.76B Demonstrating the configuration of an ultrathin device with external porous confinement.

[0105] Fig.77 mass flow rates measured from filling of an ultrathin device with 3.3 mm point spacing for human implantation with and without external porous constraints are shown according to some embodiments;

[0106] Fig.78 According to some embodiments, an entire ultrathin device (with Fig.75A H&E staining images of cell distribution within the device (similar to that shown in );

[0107] Fig.79A Showing exemplary configurations of hexagonal ultrathin devices without dots according to some embodiments;

[0108] Fig.79B Detailed view showing an exemplary configuration of a hexagonal ultrathin device without dots, according to some embodiments;

[0109] Fig.80 According to some embodiments, it is possible to fill in the blanks without dots (such as in Fig.79A in) and has a 3.3 mm dot array matrix (as in Fig.79B (middle) The amount of cells in a single ultrathin device;

[0110] Fig.81A and Fig.81BAccording to some embodiments, filling ( Fig.81A ) and filled with a porous press plate separated by 400 μm spacers ( Fig.81B ) with two configurations of a hexagonal ultrathin device with a 3.3 mm dot array matrix;

[0111] Fig.82 According to some embodiments, the present invention is presented without any constraints (such as in Fig.81A in) and in cases with porous constraints (such as in Fig.81B The amount of cells that can fill a single ultrathin device in the case of

[0112] Fig.83A An example of an ultra-thin device with a human single module design with a 2.6 mm dot pitch implanted in a minipig is shown according to some embodiments;

[0113] Fig.83B According to some embodiments, a target preperitoneal or subcutaneous implantation site in a minipig is presented at about 3 inches distal to the midline and avoiding the rib margin;

[0114] Fig.83C Subcutaneous dissection using an electrocautery in preparation for implantation of a device is shown according to some embodiments;

[0115] Fig.83D Preperitoneal dissection performed with a lighted retractor in preparation for implantation of a device is shown according to some embodiments;

[0116] Fig.84A An example of subcutaneous placement of an ultrathin device is shown according to some embodiments;

[0117] Fig.84B An example of preperitoneal placement of an ultrathin device is shown according to some embodiments;

[0118] Fig.85 Images of minipigs are shown 2 weeks after subcutaneous implantation (SQ) and preperitoneal implantation (PP) of the ultrathin device according to some embodiments;

[0119] Fig.86A An image showing an exemplary positive mold for a channel array device is shown according to some embodiments.

[0120] Fig.86B An image showing an exemplary negative mold for a channel array device is shown according to some embodiments.

[0121] Fig.87 An image showing an exemplary rodent channel array device with a manual fill tube inserted above a frame according to some embodiments;

[0122] Fig.88A Demonstrating mass flow rates of a channel array device having a fill port according to some embodiments;

[0123] Fig.88B Demonstrating mass flow rates of a channel array device with integrated fluid paths according to some embodiments;

[0124] Fig.89A An image showing exemplary geometric parameters of a channel array device according to some embodiments;

[0125] Fig.89B Illustrations showing exemplary channel test devices having a high aspect ratio (top) and a low aspect ratio (bottom), according to some embodiments;

[0126] Fig.89C A graphical representation showing cylindrical design parameters for an exemplary channel test apparatus according to some embodiments;

[0127] Fig.90A Illustration showing a high density channel array (150 μm pitch) according to some embodiments;

[0128] Fig.90B Illustration showing a low density channel array (270 μm pitch) according to some embodiments;

[0129] Fig.91A Detailed images showing an exemplary laser drilled channel array device according to some embodiments;

[0130] Fig.91B An image showing an exemplary laser drilled channel array device according to some embodiments;

[0131] Fig.91C A diagram showing a method for measuring the concentricity of an opening in a channel according to some embodiments;

[0132] Fig.92 Diagram showing measured concentricity of exemplary openings and channels

[0133] Fig.93A An image showing an exemplary laser drilled channel array device according to some embodiments;

[0134] Fig.93B An image showing a sealing interface of a channel array device according to some embodiments;

[0135] Fig.94A According to some embodiments, a high-resolution image of vascularization around an implanted channel array device is shown;

[0136] Fig.94BA low magnification image showing vascularization around an implanted channel array device according to some embodiments;

[0137] Fig.95 A diagram showing vascular host integration according to some embodiments;

[0138] Fig.96A Equalization within an exemplary channel array device is shown according to some embodiments. 2 Tension distribution;

[0139] Fig.96B Equalization within an exemplary channel array device is shown according to some embodiments. 2 Tension distribution;

[0140] Fig.97A An image showing thermoformed channels of an exemplary channel array device according to some embodiments;

[0141] Fig.97B An image showing the interior chamber height of a single channel of a channel array device according to some embodiments;

[0142] Fig.98 A graph showing measurement of fusion peel force of exemplary films according to some embodiments;

[0143] Fig.99A Images showing exemplary burst and fully formed membrane channels according to some embodiments;

[0144] Fig.99B An image showing a thermoformed film according to some embodiments;

[0145] Fig.100 showing a bar graph showing characteristic heights of exemplary films according to some embodiments;

[0146] Fig.101 showing a bar graph illustrating fusion peel forces of exemplary films formed using a fusion tool according to some embodiments;

[0147] Fig.102A a line graph showing differential scanning calorimetry analysis of an exemplary film formed using a fusion tool according to some embodiments;

[0148] Fig.102B Electron micrographs showing exemplary sintered and non-sintered films according to some embodiments;

[0149] Fig.103 Illustration showing a non-load cell enabled tool for heat fusing a membrane by spot welding according to some embodiments;

[0150] Fig.104An image showing an exemplary channel array device with inconsistent fusion fabricated using a non-load cell enabled position-based fusion tool according to some embodiments;

[0151] Fig.105A showing stress-strain curves for channel array bending with non-uniform fusion fabricated using a non-load cell enabled position-based fusion tool, wherein fusion strength decreases across the channel array device, according to some embodiments;

[0152] Fig.105B An image showing an exemplary channel array device in a peel test according to some embodiments;

[0153] Fig.106A An image showing an exemplary channel array device fabricated using a non-load cell enabled position-based fusion tool, wherein the fused areas are not aligned with the laser drilled areas, according to some embodiments;

[0154] Fig.106B An image showing an exemplary channel array device fabricated using a non-load cell enabled position-based fusion tool, wherein a leak is detected in a leak test post-fusion and the leak is repaired using an adhesive, according to some embodiments;

[0155] Fig.107 An image showing an exemplary channel array device having a single row fabricated using a load cell enabled load / force based fusion tool according to some embodiments;

[0156] Fig.108 A bar graph showing fusion strength / peel force (N) versus fusion time for a channel array device fabricated using a load / force based fusion tool enabled by a load cell is measured according to some embodiments;

[0157] Fig.109 An image showing an exemplary channel array device having an array design and a fusion strength of about 0.4N to 0.6N (left), and an image showing an exemplary channel array device having a single row design and a fusion strength of about 0.45N (right), according to some embodiments;

[0158] Fig.110 Stress-strain curves showing an exemplary channel array device having a first deformable membrane and a second flat membrane according to some embodiments;

[0159] Fig.111 Images showing exemplary channel array devices fabricated using 2X, 4X, 6X, or 8X fusion hits according to some embodiments;

[0160] Fig.112A bar graph showing the measured fusion strength / peel force (N) versus the number of fusion strikes for fusing a first deformed film to a second flat film using a load / force based fusion tool enabled by a load cell at a fusion temperature of 800°F, a fusion time of 0.05 seconds, and a fusion force of 6b according to some embodiments;

[0161] Fig.113 A bar graph showing Fusion Strength / Peel Force (N) versus Fusion Force measured for an exemplary channel array device having a first deformed film and a second flat film at a fusion temperature of 800°F, a fusion time of 0.05 seconds, and a fusion hit number of 1x according to some embodiments;

[0162] Fig.114 An image showing an exemplary channel array device having a first deformable membrane and a second flat membrane fabricated using a load-enabled fusion tool in a tensile test, wherein there is membrane drag resulting in a double fusion point, according to some embodiments;

[0163] Fig.115 A bar graph showing fusion strength / peel force (N) versus target fusion force using 4X fusion strike for an exemplary channel array device is shown according to some embodiments;

[0164] Fig.116 shows a bar graph showing Fusion Strength / Peel Force (N) of an exemplary channel array device according to some embodiments;

[0165] Fig.117A An image showing an exemplary channel array device with a 3x3 array of current points according to some embodiments herein is shown;

[0166] Fig.117B A bar graph showing fusion strength / peel force (N) of an exemplary channel array device according to some embodiments;

[0167] Fig.118 shows a bar graph showing Fusion Strength / Peel Force (N) of an exemplary channel array device according to some embodiments;

[0168] Fig.119 An image showing an exemplary channel array device in a peel test, wherein the fusion points remain intact when the film tears, according to some embodiments;

[0169] Fig.120 Showing stress-strain curves of exemplary channel array devices fabricated using non-load cell enabled or load cell enabled fusion tools according to some embodiments;

[0170] Fig.121According to some embodiments, images of fused and non-fused regions of a membrane using a loading cell enabled fusion tool are displayed;

[0171] Fig.122A An image showing channels of an exemplary channel array device fabricated using a load cell enabled fusion tool showing average seal / shelf size at fusion points according to some embodiments;

[0172] Fig.122B An image showing an exemplary channel array device fabricated using a load cell enabled fusion tool, showing concentricity between the laser drilled holes in the fusion point and the fusion area, according to some embodiments; and

[0173] Fig.123 A bar graph showing the fusion force / peel force (N) of laser-pre-channel array devices fabricated using generation 1 and generation 2 fusion tools is shown according to some embodiments. [Specific implementation method]

[0174] The present disclosure generally relates to medical devices and methods. The medical devices may include cell containment devices, devices related thereto, and methods of making and utilizing such devices. The devices may help provide improved mass transfer between environments external to and internal to the device.

[0175] In some cases, the cell-receiving device may include a high surface area to volume ratio. The high surface area to volume ratio may enable the device to provide improved mass transport into and / or out of the device, so that nutrients can be more efficiently delivered to cells within the device. In some cases, the cell-receiving device may include a first surface. The first surface may define the outer surface of the device and have a surface area. The cell-receiving device may also include a second surface opposite to the first surface, wherein the second surface defines the inner surface of the device. The cell-receiving device may also include a compartment enclosed within the second surface, wherein the compartment provides a volume for accommodating cells within the device. The device may include a single continuous open space having the volume. The first surface or the second surface of the device may include a plurality of nodes interconnected by a plurality of fibrils. The device may also include a plurality of channels running through a transverse plane of the device. The channels may provide a high surface area to volume ratio for the cell-receiving device. Each channel may include a diameter equal to or greater than 400 μm, wherein the diameter may be measured at the narrowest point in the channel. Each of the plurality of channels may be separated from each other by a distance of no more than 450 μm. The device may include a thickness greater than 250 μm measured along the transverse plane of the device. In some cases, the channel may make the thickness of the cell-receiving device not a problem for mass transport of (e.g., nutrients) into and / or out of the device. The first surface or the second surface of the device may include PVDF, PTFE, ePTFE, PCL, PE / PES, PP, PS, PMMA, PLGA or PLLA. The device may also include a frame, wherein the frame is configured to accommodate one or more cell-receiving devices. The frame may include a flexure mechanism to prevent the buckling of the cell-receiving device. The device may include a coating with a hydrophilic polymer. The volume for accommodating the cells may be inversely proportional to at least one of the diameter of the multiple channels and the number of channels per unit area of ​​the device. The surface area of ​​the device to the volume ratio may be proportional to at least one of the diameter of the multiple channels and the number of channels per unit area of ​​the device. The surface area of ​​the device to the volume ratio can achieve a larger mass transport into and / or out of the device.

[0176] In some cases, the cell containing device may include a short oxygen diffusion distance. This low oxygen diffusion distance may be independent of the size of the cell containing device, such as its thickness. For example, the cell containing device may include a substrate and a top surface relative to the substrate. A compartment for containing cells may be enclosed between the substrate and the top surface, and in some cases may rely on mass transport from outside the cell to approach nutrients to maintain survival. The height extending from the substrate to the top surface along the transverse plane of the device may have a high value, for example, greater than 300 μm. In any case, the oxygen diffusion distance of the device may be less than 150 μm. In some cases, this may be achieved by various channels or lumens of the device. In some cases, the channel may extend across the transverse plane of the device and may enable the device to have a low oxygen diffusion distance, regardless of the thickness of the device. The substrate may be substantially flat.

[0177] Two films can be used to manufacture the cell containing device described herein. In one step, various materials described herein can be used to provide a first film. In some cases, the first film can be arranged in a channel, in which the temperature and / or pressure can be set and / or adjusted to a predetermined value. The first film can be deformed (for example, to form a compartment or volume for accommodating cells). Afterwards, the second film can be fused to the first film to form the compartment. Optionally, various means such as lasers can be used to produce pores in the channel of the device to allow a continuous passage through the cell containing device. In some cases, the temperature and / or pressure can be adjusted to deform the first film or to fuse the first film to the second film. The temperature and / or pressure used in each of these processes can be an integral part of manufacturing the cell containing device. Adjusting the temperature and / or pressure of the deformation or fusion zone can control the characteristics of the feature. In some cases, increasing the temperature and / or pressure of the deformation or fusion zone can increase the depth of the feature. In some cases, after deformation, the first film can be embossed. In some cases, after fusion, the device can have a substantially planar surface and a relief surface opposite the substantially planar surface. Figure 1 A high magnification image of a cell containment device made of a PVDF membrane and having a regularly spaced array of pores within the channel of the device is shown. Deforming the membrane may also or alternatively include defibrillation, wherein nodes from one membrane unwind and interact with fibrils from another membrane, thereby creating entanglements and seals.

[0178] As described above, the device for encapsulating cells producing various bioproducts can be of great practical value, for example, for delivering treatment. The device may be referred to as a cell-containing device in this article and may include a matrix contained therein. The matrix may be a biomaterial in the inner space of the cell-containing device. The matrix may include a hydrogel, a porous sponge, an electrospun fiber, a polymeric material or other porous biocompatible materials. The matrix may also include growth factors, nutrients or other agents for enhancing cell activity and the synthesis of bioproducts. The matrix may be filled with cells or other protein expression systems, such as a cell-free expression system, to achieve the production of bioproducts. As the thickness of the matrix increases, the availability of oxygen and other nutrients may be farther and less from the edge surface of the matrix. For example, this may occur when the delivery of oxygen and nutrients mainly depends on passive delivery or diffusion. This may cause very few nutrients or no nutrients to be delivered to the region away from the surface in the matrix. The availability of nutrients may be further exhausted by cell consumption when nutrients are delivered through the matrix. For cells or systems with high demand for oxygen or other nutrients, the reduction of nutrients in the matrix may be problematic. Cell oxygen consumption can increase beyond basal state under the situation of stimulation, because cell becomes very active after described stimulation.The availability of nutrients important for cell survival rate and activity reduces and may limit the increase of the size of matrix and the cell accommodation device that surrounds described matrix.The mass transport restriction in matrix may limit the increase of device size and cannot increase the output of bio-product.It may not be desirable to reduce the density of the cell in matrix to reduce consumption in general, because it will be impractical to increase the matrix size in order to make up for the lower cell density.

[0179] The design of the cell-retaining device can be adjusted to control the surface area to volume (SA:V) ratio of the device. In some cases, a design for increasing the SA:V ratio of the current device can be provided. An increase in the SA:V ratio can help improve the delivery (for example, nutrients) to the internal area of ​​the device. One such design for increasing the SA:V ratio can be combined with channels and / or other geometries that run through the matrix and the cell-retaining device. In some cases, the channel can help increase the SA:V ratio of the device and the matrix contained in the device. The channel may include a through hole or lumen that extends completely from one side of the cell-retaining device to the opposite side. The channels may be arranged in a pattern, or may not be arranged in a distinct pattern. As an example, the channels may be arranged in an array pattern with a predetermined spacing between channels. Alternatively, the channels may be arranged in a random pattern.

[0180] Including a channel to a cell-retaining device may allow the size of the device to be increased more easily than a device without a channel. Figure 3Shown are regions with different oxygen pressures within a bilayer flat sheet and channel array device. Figure 3 As shown in , a device 300 with a channel array 301 (also referred to herein as a channel array device) can have channels extending through the matrix and the cell-receiving device. The cell-receiving device 300 with channels can have any thickness, but still avoid areas with low oxygen or nutrients. The thickness of a traditional double-layer flat sheet can be limited to 300 μm or less to avoid areas with low oxygen or nutrients. Compared to a double-layer flat sheet design or other designs without channels, a cell-receiving device with channels can have enhanced mass transport and enhanced vascularization possibilities. These features can allow the size of the channel array device to be more easily increased than a device without channels.

[0181] Cell containing devices with channels, also referred to as channel array devices, can provide an increase in surface area to allow mass transport to occur. The increase in mass transport can provide an increase in the total diffusion flux within the matrix and the device. This increase can allow matrices and devices with larger sizes to have fewer areas with little or no nutrients. Therefore, the device can have a maximum oxygen diffusion distance of less than 150 μm. Optionally, cell containing devices with channels can allow for increased vascularization possibilities. Figure 4 The surface area and vascularization potential of a device with a flat configuration and a device with channels are shown. Figure 5 Devices with flat configurations or devices with channels and increased vascularization are shown. Figure 6 Computer-aided design (CAD) renderings showing hexagonal devices with channels of increasing size but with similar SA:V ratios. Figure 4 As shown in , a device with channels can allow more cells per unit area than a device with a flat structure, because the cells can be provided with sufficient nutrients in the matrix to support their survival and activity. When implanted in a living body, the channels can allow vasculature to grow around and within the channels, compared to a device with a flat structure, in which vascularization is limited to the top and bottom surfaces of the device, as shown in Figure 4 and Figure 5As shown. In some cases, the characteristics of the channels and / or the number or density of the channels may allow for increased vascularization. For example, the diameter of the channels may be the cross-sectional distance of the channels. The diameter of the channels may be measured at its narrowest point in a transverse plane parallel to the plane of the second membrane. The channel density of the device may be the number of channels per unit area of the device, where the channels may be arranged such that there is a certain number of channels per unit area along the transverse plane of the device. The size, number, and / or density of the channels as described herein may provide increased vascularization and achieve increased mass transport of nutrients to the cells. The cell-containing device may protect the matrix and the cells or other contents of the matrix from direct contact with the vasculature in the channels. The design of the array, including its channel size and spacing, may be altered to vary the surface area and the growth pattern and likelihood of vasculature.

[0182] The dimensions of the channels can be adjusted to control the volume and the SA:V ratio within the cell-containing device. In some embodiments, the diameter of the channels can be increased to reduce the volume within the cell-containing device. Conversely, the increased diameter of the channels can provide a higher SA:V ratio for the cell-containing device. In some embodiments, the diameter of the channels can be decreased to increase the volume within the cell-containing device. The decrease in the diameter of the channels can provide a lower SA:V ratio for the cell-containing device.

[0183] The matrix can be connected as one or more pieces contained within the cell-containing device. The matrix can include a single continuous piece within the cell-containing device. The matrix can also have channels extending through its thickness at multiple locations where the channels extend through the cell-containing device. The matrix can be a biomaterial within the internal space of the cell-containing device. The matrix can include hydrogels, porous sponges, electrospun fibers, polymeric materials, or other porous biocompatible materials. The matrix can also include growth factors, nutrients, or other agents for enhancing cell activity and the synthesis of biological products. The matrix can be seeded with cells or other protein expression systems to achieve the production of biological products. Figure 7 Scanning electron micrographs showing the presentation of the cell-containing device and a cross-section of the cell-containing device with a connected interior, where the matrix can be contained within the connected interior.

[0184] The cell containing device can have various lengths, widths and heights suitable for its application. The length can be the longest dimension on the top surface of the device. The width can be the dimension perpendicular to the length on the top surface. The height of the device can also be referred to as the thickness of the device and can extend from the substrate to the top surface along the transverse plane of the device. In some cases, the length of the cell containing device can be equal to or greater than about 0.2cm, 0.5cm, 1.0cm, 1.5cm, 2.0cm, 2.5cm, 3.0cm, 4.0cm, 5.0cm, 6.0cm, 7.0cm, 8.0cm, 9.0cm, 10cm, 20cm, 30cm, 40cm, 60cm, 100cm, 120cm, 150cm, 180cm or 200cm. In some cases, the width of the cell-retaining device can be equal to or greater than about 0.2 cm, 0.5 cm, 1.0 cm, 1.5 cm, 2.0 cm, 3.0 cm, 4.0 cm, or 5.0 cm, 6.0 cm, 7.0 cm, 8.0 cm, 9.0 cm, or 10 cm. In some cases, the height of the cell-retaining device measured along the transverse plane of the device can be equal to or greater than about 100 μm, 200 μm, 300 μm, 400 μm, 500 μm, 600 μm, 700 μm, 800 μm, 900 μm, 1000 μm, 0.2 cm, 0.3 cm, 0.4 cm, 0.5 cm, 0.6 cm, 0.7 cm, 0.8 cm, 0.9 cm, 1.0 cm, 2 cm, 3 cm, 4 cm, or 5 cm.

[0185] The cell containment device can be designed to have a SA:V ratio suitable for the delivery of nutrients and desired products through the device. In some cases, the SA:V ratio can be equal to or greater than 50 cm -1 In other cases, the SA:V ratio may be equal to or greater than about 20 cm -1 、40cm -1 、60cm -1 、80cm -1 , 100cm -1 、120cm -1 、150cm -1 , 200cm -1 、250cm -1 、300cm -1 or any value therebetween. The maximum oxygen diffusion distance of the device may be less than 50 μm, 100 μm, 150 μm, 200 μm, 250 μm, 300 μm, 350 μm, 400 μm, 450 μm, or 500 μm.

[0186] In some cases, the channel 751 of the cell containment device 750 can be generally cylindrical in shape. Figure 3 , Figure 6 and Figure 7 The cylindrical channel in the cell holding device is shown. Although the channel with a cylindrical shape is mainly described herein for illustration, it should be understood that the channel can be any shape, for example, the wall of the channel can be substantially straight, curved, barrel-shaped or other shapes. In some cases, the cross-sectional area of ​​the channel can change from the top surface of the first film 711 of the cell holding device to the base of the second film 720. In some cases, the device may include a fusion portion 754. For example, the device may include a first film 711 fused to the second film 720, thereby providing a compartment 753 in the cell holding device. The compartment 753 can be filled with cells. In some cases, the fusion portion of the channel (for example, the portion where the first film meets the second film) can be substantially circular or other shapes. As described elsewhere herein, in some cases, an opening 752 can be made in the fusion portion of the film to provide a channel that runs through the cell holding device. The channel as described herein can include a cross-sectional distance or diameter. Alternatively, when the cross section of the channel is substantially circular, the cross-sectional distance of the channel can be referred to as a diameter. The diameter of the channel can be measured at its narrowest point in a transverse plane parallel to the plane of the second film. Alternatively, the diameter can be measured by the average value of the channel width along the height of the channel or device. Alternatively, the diameter can be measured at its widest point in a transverse plane parallel to the plane of the second film. In some cases, the diameter of the channel can be equal to or greater than 100 μm, 200 μm, 300 μm, 400 μm, 500 μm, 600 μm, 700 μm, 800 μm, 900 μm or 1000 μm. Optionally, the channel can have a height proportional to the diameter. In some cases, the height to diameter ratio of the channel can be equal to or greater than about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 3, 4, 5, 6, 7, 8, 9, or 10.

[0187] A plurality of channels may extend through the transverse plane of the device. The channels may be arranged so that there are a certain number of channels per unit area along the transverse plane. The number of channels per unit area may also be referred to herein as the channel density of the device. In some cases, the number of channels per unit area along the transverse plane may be equal to or greater than about 10 channels / cm 2 , 15 channels / cm 2 , 20 channels / cm 2 , 25 channels / cm 2 , 30 channels / cm 2 , 35 channels / cm 2, 40 channels / cm 2 , 45 channels / cm 2 , 50 channels / cm 2 , 60 channels / cm 2 , 70 channels / cm 2 , 80 channels / cm 2 , 90 channels / cm 2 , 100 channels / cm 2 , 110 channels / cm 2 , 120 channels / cm 2 , 130 channels / cm 2 , 140 channels / cm 2 , 150 channels / cm 2 , 175 channels / cm 2 or 200 channels / cm 2 .

[0188] The channels may be spaced apart to remove areas that receive less or no oxygen or other nutrients, which are important for cell survival and activity. In some cases, the channels may be spaced apart or separated from each other by a distance that is equal to or not greater than about 100 μm, 200 μm, 300 μm, 400 μm, 500 μm, 600 μm, 700 μm, 800 μm, 900 μm, or 1000 μm. Optionally, the distance may be measured from the center of one channel to the center of an adjacent channel. In some cases, the cell-retaining device may have a channel spacing of one distance throughout the device. Alternatively, the cell-retaining device may have channel spacings of multiple different distances across the cell-retaining device. In some cases, the channels may be arranged in a regular array having regular channel spacing distances across the device. For example, as Figure 6 As shown above, the channels can be arranged in a hexagonal array. Alternatively, other arrangements of channels can be provided, such as circular, square, etc.

[0189] In some cases, the area of ​​the lumen of the channel can be proportional to the cross-sectional area of ​​the channel. The lumen can be cut out from a portion of the fusion zone of the device. In some cases, the area of ​​the lumen can be equal to or greater than about 0, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 99% of the cross-sectional area of ​​the channel.

[0190] Multiple lumens can extend through the transverse plane of the device. The lumens can be arranged so that there is a certain number of lumens per unit area along the transverse plane. The number of lumens per unit area may also be referred to herein as the lumen density of the device. In some cases, the number of lumens per unit area along the transverse plane may be equal to or greater than about 10 lumens / cm 2 , 15 lumen / cm 2 , 20 lumen / cm 2 , 25 lumen / cm 2 , 30 lumen / cm 2 , 35 lumen / cm 2 , 40 lumen / cm 2 , 45 lumen / cm 2 , 50 lumen / cm 2 、60 lumen / cm 2 , 70 lumen / cm 2 , 80 lumen / cm 2 , 90 lumen / cm 2 , 100 lumen / cm 2 , 110 lumen / cm 2 , 120 lumen / cm 2 , 130 lumen / cm 2 , 140 lumen / cm 2 , 150 lumen / cm 2 , 175 lumen / cm 2 or 200 lumen / cm 2 .

[0191] The channel array device can be applied to various in vivo and in vitro applications. In one example, the device can contain cells or expression systems with islet cell-like functions in its matrix. The matrix can include isolated islet cells, isolated cells from the pancreas, isolated cells from tissue, stem cells, stem cell-derived cells, induced pluripotent cells, differentiated cells, transformed cells or expression systems that can synthesize one or more biological products. Optionally, the matrix can include cells of a second type that support cells of the first type, and the cells of the first type synthesize one or more biological products. The cells can be encapsulated before being placed in the matrix. The cells can be encapsulated in microcapsules or have a conformal coating. This device can be used to supplement islet cell function.

[0192] The design of the channel array in the device can affect the vascularization possibility or can run through the device growth and the amount of the blood vessel growing around the device. The channel in the device can be designed to increase the vascularization possibility. Such devices can have improved nutrients and transport and be accommodated in the lower risk of cellular hypoxia therein. Such devices can have larger device sizes, and accommodate larger matrix with cells or other expression systems. Such devices with increased transport can increase the cell survival period to more than one year after initial generation. In some cases, the cell survival period can be more than 1 month, 2 months, 4 months, 6 months, 8 months, 10 months, 12 months, 14 months, 16 months, 18 months, 20 months, 22 months, 24 months, 36 months, 48 ​​months or longer.

[0193] The change in the surface area of ​​the channel array device can affect the dynamics of the biological product. The biological product may include human cells, animal cells or transgenics. Due to the increase in surface area, the channel array device can increase the production and release of the biological product. Fig.32 The effect of device design on insulin response and surface area to volume ratio is shown. Fig.32 As shown in , the channel array device can increase the production and release of biological products as compared to a similar sized device without a channel array. The channel array device can have a higher SA:V ratio than a similar sized device without a channel array. The increase in the SA:V ratio in the channel array device can allow a higher bioproduct flux than the bioproduct flux in a similar sized device without a channel array. For a matrix with cells having islet cell-like functions, the increase in the SA:V ratio can allow a higher islet equivalent (IEQ) per unit area. In some cases, this allows increased insulin production and increased insulin flux. Figure 6 Presentation of hexagonal channel array devices of increasing size with similar SA:V ratios is shown. Fig.18 Presentation of cell containment devices with similar overall dimensions as well as variations in channel dimensions to achieve different SA:V ratios is shown.

[0194] In some cases, as further described below, the devices described herein can be assembled and / or mounted to a frame. The frame can be configured to accommodate a cell-retaining device or a plurality of cell-retaining devices. Optionally, in addition to the frame, the device can also be mounted to a subframe. The frame can provide a flexible support for the device (e.g., a channel array device) of the present disclosure. The frame can prevent unwanted folding of the device. The frame can have one or more flexure mechanisms that prevent the buckling of the cell-retaining device. The flexure mechanism can prevent the buckling of the sensitive device area in the device. The flexure mechanism can have a notch that allows the frame to bend. The flexure mechanism allows the assembly to flex along the tissue at its implantation location. Optionally, the device described herein can be utilized without a frame. For example, the device can be implanted individually in an individual without using any structural support or frame. Fig.40 Prototypes of three cell containment devices assembled onto a frame for human use are shown. The devices and the frame holding the devices can be made of materials that induce a low foreign body response. The materials of the devices and the frame can be selected to reduce inflammation or fibrosis. The devices and the frame can be used with anti-inflammatory or anti-macrophage therapies to further reduce foreign body responses.

[0195] Simple methods can be used to make cell containment devices. In some cases, the cell containment device can be made by deforming a first membrane into the shape of an array of channels and fusing a second membrane to the deformed first membrane. In some cases, a tool can be used as a guide to deform the first membrane. In some cases, a tip that takes the shape of a channel can be used to deform the membrane where the tip contacts the membrane.

[0196] The membrane may include a biocompatible porous material. The material of the membrane may allow the bioproduct to diffuse about 6 kDa or less after the manufacturing step. Alternatively, the material of the membrane may allow the bioproduct to diffuse equal to or less than about 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 200, 300, 400 or 500 kDa after the manufacturing step. The material of the membrane may have an average pore size equal to or less than about 1 nm, 2 nm, 3 nm, 4 nm, 5 nm, 6 nm, 7 nm, 8 nm, 9 nm, 10 nm, 12 nm, 14 nm, 16 nm, 18 nm or 20 nm after the manufacturing step.

[0197] One or more devices, such as film formation or fusion devices, can be used to perform the manufacturing steps of film formation, bonding and cutting. The device can be manufactured by three-dimensional printing method, micro-machining or other processing technology. In some cases, the device can be modular. In some cases, the shape of the device can affect the size and / or shape of the channel. The device can include a male mold and / or a female mold for the film (for example, the first film mentioned above). The device can be made of metal.

[0198] The device may include a platform for the film. In some cases, the platform may include a mold (e.g., a female mold) for the cell-containing device. In some cases, the platform may include a plate. The platform may also include an incision, or a hole into which the film may be molded. The incision may affect the diameter of the passage (or lumen) to be made for the cell-containing device. In some cases, the platform may be placed at a predetermined height to deviate from different surfaces. The different surfaces may be surfaces to which the film is configured to be pressed using the device described herein. This deviation height may determine the depth of the passage in the film (e.g., the first film).

[0199] The device may include various tools. In some cases, the tool may include a tip or multiple tips for deforming and / or fusing the membrane. The tool may deform the first membrane by depressing a portion of the first membrane. The tool may include: a substantially flat surface, the surface being configured to be parallel to the first membrane; and one or more protrusions on the surface, the protrusions may deform a portion of the first membrane or depress a portion of the first membrane. In some cases, the tool may include a plurality of protrusions. Each of the plurality of protrusions may include a cylinder.

[0200] A device for manufacturing a cell containing device can be configured as a support membrane (e.g., the first membrane described above). The platform can include a hole within or below the support, and the second membrane can be located in the hole. A tool including a plurality of protrusions can be configured to be connected to the platform. In some cases, the tool can be pressed down on the support platform. Optionally, the protrusion of the tool can be configured to cooperate with the hole of the platform. In some cases, the first membrane can be located on the top of the platform (e.g., above its hole). The tool can be pressed down on the first membrane and can form a deformed membrane by pressing the portion of the membrane through the hole. In some cases, the deformed membrane can be fused with the second membrane using the appropriate pressure and / or temperature described herein.

[0201] The chamber of the molding machine for holding various devices (e.g., platform and the above-mentioned tool) and film for manufacturing the cell-containing device. The chamber can be sealed. The chamber can be configured to hold the platform and / or tool. The chamber can provide a predetermined or desired temperature necessary for the occurrence of membrane fusion. The molding machine can have a sealed chamber with an air inlet and / or a vent. The molding machine can be configured to hold the film. For example, the molding machine can include a platform. A flat first film can be placed in the sealed chamber of the molding machine. Sequentially, or simultaneously, when the chamber is heated to a predetermined temperature, nitrogen or other gases can be introduced into the sealed chamber via the air inlet, while the vent is closed to reach a predetermined pressure. Using a tool, the film can be deformed, and a deformed first film can be produced. In some cases, the vent can be opened to ventilate the sealed chamber after the deformation step is completed.

[0202] As described above, a tool may be used to deform the first membrane. The tool may include a single tip or multiple tips. In some cases, a tip in the shape of a channel may be used to deform the membrane at a location where the tip contacts the membrane. The shape of the tip may be cylindrical, conical, or tapered cylindrical, or other shapes. The tip may have a contact area at the free end. The contact area of ​​the tip may contact the membrane. The area of ​​the tip may be equal to or less than about 0.5 mm 2 , 0.6mm 2 , 0.8mm 2 , 1.0mm 2 , 1.2mm 2 , 1.4mm 2 , 1.6mm 2 , 1.8mm 2 , 2.0mm 2 , 2.2mm 2 , 2.4mm 2 , 2.6mm 2 , 2.8mm 2 , 3.0mm 2 , 4.0mm 2 or 5.0mm 2 The area of ​​the tip may be equal to or greater than about 0.2 mm 2 , 0.3mm 2 , 0.4mm 2 , 0.5mm 2 , 0.6mm 2 , 0.7mm 2 , 0.8mm 2 , 0.9mm 2 , 1.0mm 2 , 1.2mm 2 , 1.4mm2 , 1.6mm 2 , 1.8mm 2 , 2.0mm 2 , 2.2mm 2 , 2.4mm 2 , 2.6mm 2 , 2.8mm 2 , 3.0mm 2 , 4.0mm 2 or 5.0mm 2 The vertical distance that the tip travels after initial contact with the membrane can help determine the height of the channel. The vertical distance that the tip travels after initial contact with the membrane can be adjusted to approximately achieve a predetermined height of the channel.

[0203] The first film can be deformed into the shape of the channel array. The first film can be deformed by thermoforming. The first film can be deformed by thermoforming based on expansion. Deforming the first film can include depressing the part of the first film with a tool. In an example, the first film can be placed in a sealed chamber of a forming machine with a mold to form a channel array. The first film can be a flat sheet before the deformation step. The forming machine with the first film can be heated to a critical predetermined temperature for deformation. Sequentially and / or simultaneously, gas can be introduced into the sealed chamber via an air inlet, while the vent is closed to reach a critical predetermined pressure and produce the first film of deformation. In some cases, pressure can be provided by pumped nitrogen. The critical pressure can be applied as a positive pressure or a negative pressure. The vent can be opened to ventilate the sealed chamber after the deformation step is completed. Figure 8 A close-up view showing the deformation of a film from a flat configuration to a molded configuration with a channel. A flat first film 810 is placed on top of a mold 820 in a sealed chamber. After the chamber is heated to a predetermined temperature and a predetermined pressure is applied to the film, the first film 811 is deformed around the mold to form a channel. In some embodiments, thermoforming enables the production of a specific internal geometry with a set height and aspect ratio to achieve and improve transplantation and vascularization.

[0204] After the first film is deformed, the second film can be fused to the first film to form a cell-containing device. The second film can be substantially flat. The second film can be placed on one side of the deformed first film. The two films can be heated to a critical temperature for fusion. When the predetermined temperature and / or pressure for fusion is reached, the two films can be compressed and / or fused together. The compression of the two films can occur at a selected position on the first film. The compression can be promoted by a mold or plate having the characteristics of taking the shape and spacing of the channel array. Heating can be performed in an oven. Alternatively, the heating can be performed with a tool having a heating element. The fused first and second films can form a fusion at the interface and form a compartment. The compartments can be interconnected, resulting in a device comprising a single continuous open space with a certain volume. The compartment can be enclosed between the substrate and the top surface of the device and can accommodate cells. Fig. 9 The fusion of the formed first film 1211 with the flat second film 1220 and the resulting device 1250 having the fused first and second films are shown.

[0205] The structure of the first and second films may be varied by manufacturing process steps. Fig.10 A cross section of a cell containing device along various parts of the cell containing device is shown, wherein in the case of material deformation or fusion, the ultrastructure of the PVDF membrane changes from being mainly nodes to being mainly elongated fibrils. The first membrane or the second membrane may include a plurality of nodes interconnected by a plurality of fibrils. During the deformation or fusion step, some of the node structures may be changed into elongated fibrils. The heat level or pressure level experienced by the membrane may affect the number of node structures. Generally, in the undeformed or unfused regions of the membrane, for example, the top surface or top of the channel, more nodes and fewer fibrils may be observed. Generally, in the regions of the membrane where deformation or fusion occurs, for example, at the bottom or middle channel of the channel, more fibrils may be observed than in the membrane where less deformation or fusion occurs. Generally, in the regions of the membrane where deformation or fusion occurs, for example, at the bottom or middle channel of the channel, fewer nodes may be observed than in the membrane where less deformation or fusion occurs. The nodes may be material storage points, and the material may be stretched into fibrils under heating and / or pressure during the deformation or fusion step. When no material is converted from the nodes in the structure to the fibrils, the membrane may break during the deformation or fusion steps. It is important to perform deformation and fusion without causing the membrane to break. The number of nodes and fibril structures and the number of node and fibril structures may be specific to the material of the membrane.

[0206] The first film and the second film may be fused without additional adhesive. The first film and the second film may be self-sealed without adhesive at a critical temperature for fusion. The fusion between the first film and the second film may provide high sealing integrity. The seam between the fused first film and the second film may be difficult to visualize, and the fused portion of the first and second films may be visible, such as Fig.11 The scanning electron micrograph shown in is presented as a continuous film. High sealing integrity can allow the cell containing device to be filled under higher pressure. Optionally, an adhesive can be placed between the first film and the second film before fusion. In some cases, the adhesive can be pressure and / or temperature sensitive. Fig.38 Tools for sealing the perimeter of the device are shown. Fig.39 Scanning electron micrograph showing the peripheral edge of a device sealed in 0.5 seconds at 345°C, where the seam between the films is difficult to discern at the ultrastructural level.

[0207] The range of critical pressure and critical temperature may be different for materials used as membranes. For example, for ePTFE, there may be a desired temperature necessary to keep the membrane in a deformed shape. In some cases, the desired temperature may be the temperature at which the material becomes sintered. In some cases, the range of critical pressure and / or critical temperature may be different for each manufacturing step. For the deformation step, the critical pressure may be less than 10psi, 20psi, 30psi, 40psi, 50psi, 60psi, 70psi, 80psi, 90psi, 100psi, 110psi, 120psi, 130psi, 140psi, 150psi, 160psi, 170psi, 180psi, 190psi, or 200psi. For the deformation step, the critical temperature may be less than 100°C, 110°C, 120°C, 130°C, 140°C, 150°C, 160°C, 170°C, 180°C, 190°C, or 200°C. For the deformation step, the critical temperature can be less than 210°C, 220°C, 230°C, 240°C, 250°C, 260°C, 270°C, 280°C, 290°C, 300°C, 310°C, 320°C, 330°C, 340°C, 350°C, 360°C, 370°C, 380°C, 390°C, 400°C, 410°C, 420°C or 430°C. For the fusion step, the critical temperature may be less than 150°C, 160°C, 170°C, 180°C, 190°C, 200°C, 210°C, 220°C, 230°C, 240°C, 250°C, 260°C, 270°C, 280°C, 290°C, 300°C, 310°C, 320°C, 330°C, 340°C, 350°C, 360°C, 370°C, 380°C, 390°C, 400°C, 410°C, 420°C or 430°C. For the fusion step, the critical pressure can be less than 10psi, 20psi, 30psi, 40psi, 50psi, 60psi, 70psi, 80psi, 90psi, 100psi, 110psi, 120psi, 130psi, 140psi, 150psi, 160psi, 170psi, 180psi, 190psi or 200psi. It may be necessary to customize the combination of critical pressure and critical temperature for each manufacturing step and material used. For each material, there may be a critical pressure and / or critical temperature range in which the material can be deformed without secondary rearrangement in the crystalline region of the polymer. Under the critical pressure and critical temperature range, the nodes in the thermoelastic material can be stretched into more fibril structures to adapt to new deformation or fusion shapes. However, outside the critical pressure and critical temperature range, the material may become crystalline and ruptured during deformation or fusion.

[0208] The film may break if it is deformed or fused outside the critical temperature and pressure range for deformation or fusion. Selecting a temperature and pressure outside the critical temperature and pressure range for deformation may result in a first film that may not fuse or poorly fuse to the second film in the fusion step. This rupture in the first film may not be obvious before the fusion step. When outside the critical temperature or pressure range, the crystallinity of the material of the film may increase. The material of the first film with increased crystallinity may not fuse or fuse to the material of the second film poorly. The relative crystallinity can be measured by calculating the transition enthalpy of the material using a differential scanning calorimeter (DSC). The secondary peak of the DSC heat flow measurement of the film can indicate the rearrangement of the crystalline structure of the film and can indicate that the film may not easily fuse with another film. This rearrangement may occur when the relative crystallinity does not increase. The rearrangement may also occur when the relative crystallinity does not decrease. The arrangement of the crystalline regions of the film is an important factor in fusion, and the potential rearrangement of the crystalline structure may not allow the chain entanglement of the crystalline region to another film during fusion. Fig.15 Another example of DSC heat flow measurement of a film deformed at 100 psi and 160°C is shown, which maintains a shoulder peak compared to the baseline PVDF film, indicating that its crystalline structure does not change significantly and will fuse to a second film. Heat flow measurement of a film deformed at 100 psi and 173°C with quench cooling has a different secondary peak in the first melting endotherm, indicating that this film forms a secondary crystalline region and will not fuse to another film.

[0209] In some cases, a tip can be used to deform the first film. The tip can travel a predetermined vertical distance to roughly reach the predetermined height of the channel after the initial contact with the film. In some cases, a tip can be used to fuse two films at the position where the tip contacts the film, wherein the first film has been deformed before. Fusion can be performed with a tip, wherein the tip presses the first film and the second film to contact each other for a predetermined time. In some cases, a tip can be used in a single process or step to deform the first film and fuse the first film to the second film. In some cases, the first film deviates vertically from the second film at a predetermined height. This vertical deviation can determine the channel height. Deformation and fusion can be performed in one step with the tip, wherein the tip contacts the first film, deviates from the first film and moves vertically toward the second film and presses the first film and the second film to contact each other for a predetermined time. In some cases, a single tip can be used for the film. Fig.34 Display through 2mm 2 Schematic diagram of the fusion process whereby a tip spot welds two membranes. In other cases, multiple tips may be used on the membranes, for example, simultaneously.

[0210] In some cases, when the one or more films are fixed, the tip can move laterally (xy direction) and vertically (z direction). In some cases, when the stage holding the one or more films moves laterally, the tip can only move vertically. In some cases, the tip can travel a predetermined lateral distance relative to the film surface and then travel a predetermined vertical distance downward to enter the film and return to a neutral vertical position upward, which can be its previous vertical position. This cycle can be repeated until a predetermined number of channels are deformed and / or fused on the one or more films. In some cases, the stage holding the film can travel laterally to a position at a predetermined position below the tip, and the tip can travel a predetermined vertical distance downward to enter the film and return to a neutral vertical position upward. This cycle can be repeated until a predetermined number of channels are deformed and / or fused on the one or more films. The movement of the tip and / or the stage holding the film can be programmed and automated. The tip may travel about 100 μm, 200 μm, 300 μm, 400 μm, 500 μm, 600 μm, 700 μm, 800 μm, 900 μm, or 1000 μm laterally in the x and / or y direction between cycles. The tip may travel about 100 μm, 200 μm, 300 μm, 400 μm, 500 μm, 600 μm, 700 μm, 800 μm, 900 μm, or 1000 μm vertically after contacting the membrane.

[0211] In the process of using the tip to manufacture the cell containing device, there may be a critical time, temperature and pressure range for deformation and / or fusion of the film. The time when the tip contacts the film can be referred to as the tip contact time. In some cases, the tip contact time can be about 0.1 seconds, 0.2 seconds, 0.3 seconds, 0.4 seconds, 0.5 seconds, 0.6 seconds, 0.7 seconds, 0.8 seconds, 0.9 seconds or 1 second. The tip contact time can be about 2 seconds, 3 seconds, 4 seconds, 5 seconds, 6 seconds, 7 seconds, 8 seconds, 9 seconds, 10 seconds, 11 seconds, 12 seconds, 13 seconds, 14 seconds, 15 seconds, 16 seconds, 17 seconds, 18 seconds, 19 seconds, 20 seconds, 25 seconds or 30 seconds. The tip can be heated to a temperature higher than the critical temperature for deformation and / or fusion. Basically as described above, the tip can be heated to a temperature that is roughly the critical temperature for deformation and / or fusion. The tip can be heated to about 0° C., 1° C., 2° C., 3° ​​C., 4° C., 5° C., 6° C., 7° C., 8° C., 9° C., 10° C., 15° C., 20° C. above the critical temperature. The tip can apply a pressure of less than 10 psi, 20 psi, 30 psi, 40 psi, 50 psi, 60 psi, 70 psi, 80 psi, 90 psi, 100 psi, 110 psi, 120 psi, 130 psi, 140 psi, 150 psi, 160 psi, 170 psi, 180 psi, 190 psi, or 200 psi to the membrane for deformation and / or fusion.

[0212] In some cases, the first film can be sintered or non-sintered before fusion. In some cases, the second film can be sintered or non-sintered before fusion. In one example, before fusion, the first film can be sintered and the second film can be non-sintered. The films can be sintered at various temperatures and for various times. The sintered film can have a lower melting temperature than a non-sintered film of the same type. In one example, the first film can be sintered at 370°C for 7 minutes. Fig.35 DSC measurements of an ePTFE membrane sintered at 370°C for 7 minutes and a non-sintered ePTFE membrane at 300°C are shown, wherein the sintered membrane has a lower melting temperature of 320°C to 325°C compared to 340°C to 350°C for the non-sintered ePTFE membrane. Optionally, whether the membrane is sintered may be important for the fusion of the first membrane to the second membrane. In some cases, the membrane being sintered may help the membrane to deform and maintain the deformed shape. Optionally, the fusion of the sintered membrane to the non-sintered membrane may provide desired properties, such as tight sealing, device integrity and / or shape.

[0213] The edges of the fused first and second membranes can be trimmed to remove excess membrane on the periphery of the cell containment device. In some cases, the trimming can be performed by punching. An alignment frame can be used to mount and align the cell containment device, and a perimeter punch can be used to cut excess periphery of the cell containment device.

[0214] according to Fig. 20 The fused portion between the first and second membranes can be cut to form the lumen of the channel. The fused portion of the first and second membranes can be removed via laser ablation, thereby forming a channel traversing the device. Fig.21 Scanning electron micrograph and image showing a cell containment device having a lumen 2752 with a fused portion 2754 cut by laser ablation and a cell containment device with a frame. Fig. 22 A scanning electron micrograph of a cross section of a cell containment device is shown after a manufacturing step including cutting through the channel to form a lumen. The cutting may be performed by laser etching or laser ablation. The removed portion may be a portion of the area of ​​the fused portion so as not to compromise the seal between the first membrane and the second membrane. The removed portion may be about 0, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 99% of the area of ​​the fused portion.

[0215] The assembled cell containment device may be provided as part of a modular system, referred to herein as a macro device. Fig.23 One or more cell-receiving devices 2950 are shown, each of which can be filled with a matrix including cells inside, and the cell-receiving devices can be assembled on a frame 2980 to form the macro device. Multiple channel array devices or cell-receiving devices with a matrix can be placed on the frame in various configurations. Each device can have a filling port 2955 to fill the interior of the device with a matrix. The filling port is designed to be smaller to reduce the possibility of device seal rupture. The filling port can be used to fill cells in the device. In some cases, the frame can include a flexible backing material or structural support. Optionally, in addition to the frame, the channel array device can be mounted on a subframe 2981. The frame can provide a flexible support for the channel array device. The frame can prevent unwanted folding of the device. The frame can have one or more flexure mechanisms to prevent buckling of sensitive device areas. The flexure mechanism 2982 can have a notch that allows the frame to bend at the notch. The flexure mechanism allows the assembly to flex along the tissue at the location where the assembly is implanted. The frame can have a manipulation tab 2956. The manipulation tabs may be used for surgical manipulation and / or implantation of the assembled device.The frame may have holes to allow for delivery.

[0216] The frame can have a flexure mechanism to prevent buckling of the cell-receiving device mounted on the frame. The flexure mechanism includes a small notch or cutout while keeping a small portion of the frame intact. In other cases, the notch can be in approximately the same position on the top and bottom surfaces of the frame while keeping a small portion of the frame intact between the two notches. In other cases, the notch can be on one of the surfaces of the frame while keeping a small portion of the frame intact. The notch can have various shapes, including generally conical, cylindrical, pyramidal, rectangular or other shapes that remove a portion of the frame. The notch allows the frame to bend at various angles, which can be from 0° to 90° in any given direction.

[0217] The macro device may have several configurations and sizes depending on its application. In some cases, the macro device may have a width of at least 3 cm, 4 cm, 5 cm, 6 cm, or 7 cm. In some cases, the macro device may have a length of at least 5 cm, 6 cm, 7 cm, 8 cm, 9 cm, 10 cm, 11 cm, 12 cm, 13 cm, 14 cm, or 15 cm.

[0218] The cell-receiving device may be filled with a matrix. In some cases, the cell-receiving device may be filled by pressure. In other cases, the cell-receiving device may be filled by centrifugation. Fig.24 Show the filling of framed (left) and frameless (right) PVDF cell-containing devices with matrix containing cells. In some cases, a filling tube can be connected to the cell-containing device to fill the frameless device. In other cases, in a framed cell-containing device, the frame can allow a fluid path starting from the filling hub to be connected to a filling port on the device. Once the device is filled, the device can be separated and sealed from the filling tube or filling hub. The frame can also be sealed. The device and / or the frame can be sealed with glue or UV curable glue. The matrix can include cells or expression systems that produce biological products. The matrix can also include cell culture media. The matrix can also include porous biocompatible materials. The risk of seal rupture can be reduced by leaving only a small area in the cell-containing device unsealed for filling. The small area can be sealed after filling the cell-containing device with the matrix. Fig.25 A cell-containing device filled with a matrix having cells, as visualized by a tissue section stained with hematoxylin and eosin (H&E), is shown. In some cases, the cell-containing device can be filled (e.g., filled with cells) using the following process. First, the cells can be suspended in a medium. Second, the cells can be applied to the filling port under pressure. Third, the filling port can be removed. Finally, the filling port opening can be sealed (e.g., with UV curing glue).

[0219] The surface of the assembled channel array device or its components can be treated. The surface can be treated with a material to promote vascularization. The coating can be VEGF, or other angiogenic factors or substances. The outer surface can be treated with a material that imparts antifouling properties. The surface of the device can be treated to reduce the possibility of fibrosis or connective tissue formation around the device. The material of the device can be selected to reduce the possibility of fibrosis. The surface of the device can be treated to produce physical characteristics, or the surface of the device can be treated chemically to reduce fibrosis. The surface can be treated with a hydrophilic coating. The hydrophilic coating can include polymers, polyethylene glycol, polyvinyl alcohol, polydopamine, oact. Fig.41 An example of a scheme for forming a hydrophilic coating on the surface of a membrane is shown. The hydrophilic coating can impart hydrophilic properties to a hydrophobic surface prior to coating. Improving the hydrophilicity of the membrane surface can improve the transport of hydrophilic molecules across the membrane between the internal environment and the external environment of the device. Fig.38 An ePTFE cell containment device is shown after hydrophilic coating treatment in water, with air bubbles indicating the ability of water to fill the interior of the device and displace air from the interior of the device.

[0220] The membrane of the cell containing device may include one or more porous materials. The membrane may include PTFE, ePTFE, PVDF, PCL, PE / PES, PP, PS, PMMA, PLGA, PLLA or other thermoelastic materials. The material of the membrane may be synthesized by various methods. The synthesis method of the porous material may include expansion, solvent casting, immersion precipitation and phase separation, electrospinning, a method for obtaining a homomesh network, a method for obtaining a beam-cable network or other methods. The membrane may be a porous material that allows a material with a molecular weight less than about 3000kDa, 2000kDa, 1000kDa, 500kDa, 400kDa, 300kDa, 200kDa, 100kDa, 50kDa, 40kDa, 30kDa, 20kDa, 10kDa, 6kDa, 5kDa, 4kDa, 3kDa, 2kDa, 1kDa to be transported through the material after the manufacturing process. The membrane may have an average pore size of about 5 nm, 10 nm, 15 nm, 20 nm, 30 nm, 40 nm, 50 nm, 60 nm, 70 nm, 80 nm, 90 nm, 100 nm, 200 nm, 300 nm, 400 nm, 500 nm, 600 nm, 700 nm, 800 nm, 900 nm, 1000 nm, 1100 nm, 1200 nm, 1300 nm, 1400 nm, 1500 nm, 1700 nm, 2000 nm or 2500 nm.

[0221] The dimensions of the channel array can be controlled by parameters in the manufacturing steps. A mold or plate can be used to deform the first membrane into a predetermined channel array design. The temperature during the deformation step can be used to change the depth of the channel. The pressure during the deformation step can be used to change the depth of the channel. The depth of the channel can be changed using a combination of temperature and pressure during the deformation step. The temperature and pressure during the deformation may be specific to the material of the first membrane. Changes in channel size may affect the three-dimensional shape of the interconnected recesses within the cell-receiving device. These manufacturing parameters can be used to adjust the configuration and size of cell containment of the channel array device. These parameters can be used to change the SA:V ratio of cell containment. The SA:V ratio of cell containment can improve vascularization in and around the device.

[0222] The device can be implanted in vivo at various sites in the subject. In some cases, the device on the frame can be implanted in the subject. In one example, the device can be placed by implantation before the peritoneum or after the rectus muscle. In other examples, the device can be placed by implantation within the omentum. In another example, the device can be placed by subcutaneous implantation. In another example, the device can be placed by implantation above the liver. Fig.28 Preperitoneal, intraomental, suprahepatic, and subcutaneous implantation of cell-containing devices in rats is demonstrated.

[0223] The device can be fixed in vivo at the implantation site. In one example, a tissue adhesive can be used to fix the device. The tissue adhesive can be a fibrin, cyanoacrylate, polyethylene glycol, albumin-based adhesive or a polymer-based adhesive. In another example, platelet-rich plasma can be used to fix the device.

[0224] Following implantation in vivo, blood vessels may form around the device and within channels within the device. Fig.31 and Fig.32 The vasculature surrounding the cell containment device and in the channels is shown 20 and 90 days after implantation in the preperitoneal site in rats. Fig.29 H&E stained tissue sections showing vascularization around the cell containment device and vasculature in the channels after in vivo implantation in rats. Fig.30 Vascularization around a cell-containing device with low and high channel density after in vivo implantation is demonstrated. The blood vessels may have smooth muscle cells that are typically found in arteries. The blood vessels may have arterial characteristics. Fig.30 and Fig.31The density of channels in a channel array device may affect the degree of vascularization. A higher channel density may increase the degree of vascularization. A lower channel density may decrease the degree of vascularization. The diameter of channels in a channel array device may affect the degree of vascularization and the branching of blood vessels.

[0225] The cell containment device can be designed to achieve various functional goals. One of the goals of the cell containment device can be to provide at least one year of cell survival for the cells in the device after implantation in vivo. In some embodiments, the device is designed to allow at least 4x10 8 The cells have a cell survival period of at least one year. The device can be retrieved or removed from the in vivo implantation site in the subject to evaluate the cell survival rate and other functional assessments after implantation. The cell containing device can be designed to improve the mass transport within the device and through the membrane of the device. The cell containing device can be coated and / or implanted at the site to improve the proximity to the host vascular supply. The cell containing device can be designed to stabilize the host-device interface to allow the flexibility of the device without folding on itself. The cell containing device can be designed to alleviate tissue integration to provide stability to the device without compromising the integrity of the device. The cell containing device can be designed to use materials and coatings that provide favorable non-specific biomaterial reactions.

[0226] Ultra-thin device

[0227] Although cell-containing devices including channels are primarily described herein, cell-containing devices do not necessarily need to include channels. For example, in some cases, another approach to meeting various functional goals of cell-containing devices can be met using ultrathin cell-containing devices (also referred to herein as ultrathin devices). Therefore, various parameters, properties, or descriptions (e.g., coatings, materials, etc.) described for a given embodiment of a cell-containing device (e.g., a cell-containing device including a channel) can be equally applicable to another embodiment of a cell-containing device (e.g., an ultrathin device).

[0228] The ultrathin device can have a thin total cross-sectional thickness. Alternatively or additionally, the ultrathin device can have a thin membrane. The membrane of the ultrathin device can be a biocompatible polymer or biomaterial, such as ePTFE, PVDF, PEEK, PS, PES, PAN / PVC, nylon, polyurethane, polycarbonate, polyacrylonitrile, fiberglass, polycaprolactone, hydrogel, polyester, polyanhydride or cellulose. The membrane can also be made of a permanent, non-degradable material, or alternatively, made of a biodegradable material with a controlled degradation profile. The size of the ultrathin device can provide a high SA:V ratio. A high SA:V ratio can enhance the transport of molecules into and out of the device, such as transporting nutrients and oxygen into the device and transporting insulin or other secretory products out of the device for the indwelling cells in the device. Modeled insulin diffusion outside such ultrathin devices containing insulin-producing cells can be 0.4-10ng / cm 2 / 10 minutes. The ultra-thin device may not have channels extending through the thickness of the device as with a channel array device. Figure 5 and Fig.46 Schematic diagrams of an ultrathin device (plane (flat device)) with a cross-sectional thickness of 250 μm and filled with cells (black circles) and a macro device with three ultrathin devices are shown, respectively.

[0229] The ultrathin device may have a total cross-sectional thickness of about 250 μm. In some embodiments, the total cross-sectional thickness of the ultrathin device may be less than 5000 μm, 4000 μm, 3000 μm, 2000 μm, 1000 μm, 900 μm, 800 μm, 700 μm, 600 μm, 500 μm, 400 μm, 300 μm, 200 μm, 100 μm, or 50 μm. In some embodiments, the total cross-sectional thickness of the ultrathin device may be at least 1000 μm, 900 μm, 800 μm, 700 μm, 600 μm, 500 μm, 400 μm, 300 μm, 200 μm, 100 μm, 50 μm, or 10 μm.

[0230] The ultrathin device may have a film with a thickness of 2 μm to 25 μm. In some embodiments, the film of the ultrathin device is less than 100 μm, 90 μm, 80 μm, 70 μm, 60 μm, 50 μm, 40 μm, 30 μm, 20 μm, 10 μm, 5 μm or 1 μm.

[0231] The ultrathin device can be designed to have a SA:V ratio suitable for the transport of nutrients and desired products through the device. In some cases, the SA:V ratio can be equal to or greater than 80 cm-1. In other cases, the SA:V ratio can be equal to or greater than about 20 cm-1. -1、40cm -1 、60cm -1 、80cm -1 , 100cm -1 、120cm -1 、150cm -1 or any value therebetween. The maximum oxygen diffusion distance of the device may be less than 50 μm, 100 μm, 150 μm, 200 μm, 250 μm, 300 μm, 350 μm, 400 μm, 450 μm, or 500 μm.

[0232] The properties of the membrane of the ultra-thin device can be selected to enhance its function. One such property can be the flux selectivity of the membrane. Various membrane properties, including microstructure, tortuosity, pore size, porosity and / or thickness, may contribute to the flux selectivity. The flux selectivity can affect the molecules that can pass through the membrane. Because the microstructure affects the flux selectivity of the membrane, the processing of the membrane may affect its microstructure and its flux selectivity.

[0233] The property of the film of the ultrathin device can be selected to improve its ability for immunoprotection. For example, a film with high flux selectivity can prevent a large amount of antibodies and supplementary proteins from moving through the film. The film with high flux selectivity can also reduce nutrients to diffuse to the inside of the cell accommodating device through the film. The film with medium flux selectivity can have good flux properties, to realize cell survival, reduce antigen release from dead cells and prevent antibodies from moving through the film to a certain extent. The film with low flux selectivity can allow high flux to promote the exchange of nutrients through the film and can limit the transport of cells through the film and not transport molecules smaller than cells. Equally, the film with medium or low flux selectivity can allow the potential exchange of antibodies and can have a mechanical property lower than the film with higher flux selectivity.

[0234] Various types of films can be used in ultrathin devices. Fig.47 Scanning electron micrographs of ePTFE membranes with different pore thicknesses that can be used for ultrathin devices are shown. Table 1 shows various membrane properties, including its flux, thickness, diffusion selectivity and insulin flux. The diffusion selectivity is defined as the ratio of antibody flux to insulin flux, where a lower ratio indicates a more selective flux, because insulin is a molecule much smaller than an antibody. The C membrane has a lower diffusion selectivity than the A and B membranes and is more selective in allowing larger molecules (such as antibodies) to be transported through its membrane.

[0235] [Table 1: Films for ultra-thin devices]

[0236]

[0237] like Fig.48As shown in, a high-throughput ultrathin device including a selectively permeable membrane and filled with rat islet cells can produce and secrete insulin in response to 20mM glucose stimulation, as measured by the flux of C-peptide to the outside of the ultrathin device. C-peptide, also known as a connecting peptide, is a polypeptide that splits from proinsulin to form an insulin molecule. The level of C-peptide provides the level of insulin produced and secreted, because C-peptide is present in equimolar amounts with insulin. The islets obtained from Sprague-Dawley rats are used to assess the insulin delivery dynamics after being encapsulated in the ultrathin device. The ultrathin device filled with encapsulated rat islet cells exhibits a delay of about 5 minutes to 10 minutes in the release of insulin from the ultrathin device compared to the ultrathin device filled with free rat islets, which is measured according to the dynamic GSIS (glucose-stimulated insulin secretion) over a period of time. Encapsulated islet cells can respond to glucose stimulation and produce and release insulin. Encapsulation does not hinder the ability of the encapsulated cells to receive glucose stimulation and respond to glucose stimulation by producing and secreting insulin. The ultrathin device filled with islet cells has diffusion dynamics that allow the biphasic insulin secretion and interruption that occur in response to the glucose concentration in its environment to be distinguished. When cells are exposed to glucose and secrete insulin, GSIS occurs. The amount of secreted insulin can be proportional to the level of glucose exposure. During GSIS, insulin secretion may be reduced, stopped, started and increased according to the level of glucose exposure. Some aspects also include releasing insulin from the cells in the cell-containing device in an amount that is enough to reduce the blood sugar level in the subject. In some aspects, the release of insulin stops when the blood sugar level in the subject is reduced to normal levels. In some aspects, the release of insulin restarts when the cells in the cell-containing device are exposed to high blood sugar levels in the subject again.

[0238] The choice of membrane may affect the flux kinetics of insulin from the ultrathin device as well as the responsiveness of the ultrathin device to glucose stimulation. Fig.49 Static C-peptide release from ultrathin devices including high-flux membranes or selectively permeable membranes filled with encapsulated insulin cells before (LG or low glucose), during (HG), and after (LG) high glucose stimulation is demonstrated. Encapsulated islet cells in all three types of ultrathin membranes responded to glucose stimulation. Ultrathin devices with high-flux membranes resulted in approximately 1.7 x 10 5 pM C peptide, while the ultrathin device with a selectively permeable membrane resulted in approximately 5x10 per device under HG conditions 4 pM C peptide. For the ultrathin device, there was a delay in the release of insulin in response to HG conditions. The delay in insulin release may be attributed to the retention of insulin within the ultrathin device and may indicate that some membranes may be trapping insulin. Fig.50Dynamic GSIS of an ultrathin device with AS-1 membrane is demonstrated. Since AS-1 is a high-flux membrane, this results in a high-flux ultrathin device that responds to a high glucose stimulus of 20 mM by producing and secreting insulin and stops insulin production and secretion when the high glucose stimulus is removed.

[0239] Ultrathin devices and cell-containing devices can be prepared by many methods. These methods may include film making, film coating, device assembly and aseptic cell filling. The film can be made and processed to achieve target properties. The target properties may include transport flux properties, mechanical properties, porosity, pore size, thickness, microstructure or tortuosity. The production method may include film stretching or sintering. The film can also be treated with a coating method to impart various desired properties. As described above, these coatings may include hydrophilic polymers, VEGF or other molecules to promote vascularization or protein delivery. Subsequently, a film can be made and assembled into the device. The ultrathin device can be assembled using a robotic assembly, which can be an automatic or semi-automatic assembly. A frame for the device can be made. Optionally, one or more ultrathin devices can be assembled onto the frame as a macro device. The ultrathin device can be aseptically filled with cells. The aseptic cell filling can be carried out by a pressure filling method, centrifugation, gravity filling, opening filling or any combination thereof.

[0240] The membrane can be sintered before being fabricated into the cell containment device. The sintering process of the membrane can be used to change the porosity and flux properties of the membrane. Sintering can increase the porosity of the membrane while maintaining its pore structure. Sintering can improve the mechanical stability and insulin flux of the membrane. Fig.51 Scanning electron micrographs of non-sintered and sintered films are shown and illustrate changes in the film's microstructure, where there may be fusion and coalescence of nodes and fibrils of the film in the case of sintering.

[0241] The sintering process can be very consistent with low batch-to-batch variation. Table 2 shows the melting point temperature of the sintered membrane as measured by DSC, which is about 326°C to 333°C and is different from the melting point temperature of the non-sintered membrane of about 345°C. The sintering process resulted in a batch-to-batch variation of 0.76%, indicating consistency with the sintering process. Sintering of the membrane can be used to modify the porosity of the membrane, which in turn can be used to adjust the porosity and flux properties of the cell containment device. Consistency in the sintering process can provide an attractive option for affecting membrane porosity and properties in large-scale manufacturing of cell containment devices.

[0242] [Table 2: Melting temperature of sintered films as measured by DSC]

[0243] membrane <![CDATA[T m [℃]]]> AU reference (non-sintered) 345.1 AS Round 1 326.35 AS Round 2 329.3 AS Round 3 329.15 AS Round 4 333.24 AS Round 5 330.76

[0244] In some embodiments, the cell containment device assembled to the frame can be made with a non-sintered film and then undergo post-heat curing to reduce porosity on the frame. This manufacturing method using a non-sintered film and post-heat curing for assembly can reduce steps, time and / or cost in the overall manufacturing process.

[0245] In some embodiments, the cell containing device may include at least one sintered membrane and at least one non-sintered membrane. Such asymmetric sintering of the membrane of the cell containing device can provide a device with a controlled geometry. Using different types of membranes in a cell containing device may cause bending in the device due to their different mechanical properties, wherein the first membrane may be more flexible or ductile than the second membrane. In some embodiments, the different types of membranes in a device can be sintered membranes and non-sintered membranes.

[0246] Coating of the membrane provides another method of adjusting the flux characteristics of the cell containment device. The membrane can be coated with a hydrophilic coating before being fabricated into the cell containment device. The hydrophilic coating can allow the membrane to wet, thereby allowing ultrafiltration, which can be used to load cells into the device, achieve diffusion, and provide a neutrally charged surface for biocompatibility. Fig.52 The display results in a value close to 1x10 -6 mol / m 2 Comparison of hydrophilic coating methods for membranes with a target insulin flux of 1000 ng / s. The hydrophilic coating method resulted in an intra-batch variation of about 9% and an inter-batch variation of about 8%.

[0247] In some embodiments, hydrophobic membrane can be coated with hydrophilic coating.Described hydrophilic coating can be biocompatible and can improve the diffusion of insulin and other molecules.In some embodiments, the hydrophobic membrane without coating may not allow the diffusion of insulin and other molecules.In some embodiments, nanometer thin coating method can provide permeability and insulin diffusion of appropriate level for film and cell accommodating device.In some embodiments, the semi-permeability of film is configured to protect cells from immune attack.In some embodiments, the semi-permeability of described film is configured to protect described cells from immune attack when lacking immunosuppressive therapy.

[0248] Due to the hydrophobic nature of the ePTFE material, hydrophilic polymers can be polymerized around the ePTFE microstructures to enable membrane wetting and reduce hydraulic resistance. This can facilitate ultrafiltration during cell loading and enable the use of lower pressures to introduce cells into the device. This can also create a neutral hydrophilic surface to minimize absorption or attachment of host proteins and cells.

[0249] The framed and sintered membrane can be placed in 100% ethanol for 5 minutes and then soaked in 30% ethanol for about 5 minutes. The membrane can then be soaked in a coating solution at room temperature for about 5 minutes, the coating solution containing 9g APS, 27mL HPA and 18mL TEGDA in 30% ethanol. The polymerization reaction can be carried out at 70°C with a rate of 3°C / minute from room temperature, and the temperature rise is controlled using LabView software. The framed coated membrane is removed from the coating solution and transferred to boiling 100% ethanol to remove unreacted monomers and then soaked in excess distilled water with several changes. Finally, the coated membrane is dried in a chamber with a continuous stream of nitrogen.

[0250] Fig.53 Films coated by three different coating methods V1, V2 and V3 are shown, which have been stained with H&E dye as an indication of the hydrophilicity of the film. The film subjected to the V1 coating method appears to be stained very dark pink and appears to be over-coated. The film subjected to the V2 coating method has variable H&E dye staining levels that are stronger near its edges than in the center and appears to have a gradient coating. The film subjected to the V3 coating method has been coated by the nano-thin coating method and appears to be uniformly coated over its cross-sectional thickness. Fig.54 The hydraulic permeability of the membrane, the membrane subjected to the V1 coating process, and the membrane subjected to the V3 coating process are shown, wherein the V3 coated membrane exhibits a hydraulic permeability of about 1.8x10 -14 m 2 The high permeability of the membrane and the V1-coated membrane has less than 1x10 -15 m 2 Lower hydraulic permeability.

[0251] In some embodiments, the coated film in the ultrathin device may have a thickness of at least 1×10 -16 m 2 , 1x10 -15 m 2 , 1x10 -14 m 2 or 1x10 -13 m 2In some embodiments, the first film and the second film of the ultrathin device may have the same hydraulic permeability. In some embodiments, the first film and the second film of the ultrathin device may have different hydraulic permeabilities. In some embodiments, different coating methods can be used to achieve different hydraulic permeabilities for the first film and the second film. In some embodiments, the semi-permeability of the first film, the second film, or both is configured to protect the cells from immune attack. In some embodiments, the semi-permeability of the first film, the second film, or both is configured to protect the cells from immune attack in the absence of immunosuppressive therapy.

[0252] The coating method can be designed and scaled up to coat multiple membranes or multiple cell containment devices at one time. In some embodiments, the coating method can be scaled up to coat 40 human cell containment devices at one time. Fig.55 The setup for the extended coating process and the dry and wet coated films are shown. The high light transmittance observed by transillumination of the wet coated film indicates the wettability of the coated film and serves as an indicator of the hydrophilic nature and permeability of the coated film.

[0253] The frame for the cell containment device can include various materials. In some embodiments, the frame can be a biocompatible material. As described above, the frame can hold a cell containment device or multiple cell containment devices. The frame can have mechanical properties similar to the host biological tissue surrounding the device after implantation in the body. One measure of the mechanical properties of a material is its Young's modulus. The device can be formed to show the Young's modulus of various biological materials and synthetic materials and a range of target Young's moduli for candidate membranes, candidate frame materials, and the entire device (composite Young's modulus). The target Young's modulus of the membrane can be 10 6 Pa to 10 9 Pa. The target Young's modulus of the frame material can be 10 8 Pa to 10 9 Pa. The target Young's modulus of the device composition (also referred to as a macro device or a device with a frame) can be 10 7 Pa to 10 9 Pa, between the range of the device alone and the frame alone. In some embodiments, the membrane may have at least 10 5 Pa, 10 6 Pa, 10 7 Pa, 10 8 Pa or 10 9 In some embodiments, the frame may have a Young's modulus of at least 10 7 Pa, 10 8 Pa or 10 9In some embodiments, the device composition may have a Young's modulus of at least 10 7 Pa, 10 8 Pa or 10 9 Young's modulus in Pa.

[0254] In some embodiments, the frame can include polyetheretherketone (PEEK). Fig.56 Simulations of the mechanical properties of a 300 μm thick PEEK frame for two cell containment devices under a force of 150 mN are shown. The simulations indicate that the PEEK frame will have a maximum stress of about 88 MPa (which is below the yield stress of about 103 MPa), a maximum strain of about 0.014, and a maximum displacement of about 4.711 mm.

[0255] Fig.57 An example of a PEEK frame holding a cell containment device is shown with a fusion point in the center of the device. The frame can be micromachined or machined using methods appropriate to achieve the target dimensions of the frame. Fig.58 A single frame module is shown, comprising a single cell containment device on the frame with a centered fusion point, the cell containment device being maximally filled. The filled device with a centered fusion point on the frame shows limited lateral expansion of the membrane.

[0256] Fig.75A and Fig.75B Various configurations of macro devices with multiple devices held by a macro device frame are shown. In one design, the macro device frame can be flexible and hold multiple devices. In some embodiments, the macro device frame can be a flexible integrated frame for holding multiple devices and has a porous structure around a single cell containing device.

[0257] like Fig.59 As shown in , the cell receiving device can be filled with cells to the greatest extent. Fig.60Demonstrate cell viability of at least 10 days in maximally filled cell containment devices and stored under standard conditions of 20% oxygen and 37°C, as evidenced by the presence of cells in H&E stained tissue sections. Cell containment devices filled with cells can be stored under various conditions to extend the viability of cells within the cell containment devices prior to implantation. Cell containment devices filled with cells can be stored at various temperatures, either 4°C, 23°C, or 37°C. In some embodiments, the cell-holding devices filled with cells can be stored at a temperature of at least 1° C., 2° C., 3° ​​C., 4° C., 5° C., 6° C., 7° C., 8° C., 9° C., 10° C., 11° C., 12° C., 13° C., 14° C., 15° C., 16° C., 17° C., 18° C., 19° C., 20° C., 21° C., 22° C., 23° C., 24° C., 25° C., 26° C., 27° C., 28° C., 29° C., 30° C., 31° C., 32° C., 33° C., 34° C., 35° C., 36° C., 37° C., 38° C., 39° C., or 40° C. In some embodiments, the cell-holding devices filled with cells can be stored under hypoxic, normoxic, or hyperoxic conditions.

[0258] The two membranes of the cell containment device can be fused into discrete points along its surface. Various configurations of fusion point shapes, diameters or distances and densities (e.g., center-to-center spacing) of the device can exist. The points can be circular, rectangular, triangular, linear or other shapes. The points can have various cross-sectional distances or diameters. In some embodiments, the dot diameter can be at least 0.1 mm, 0.2 mm, 0.3 mm, 0.4 mm, 0.5 mm, 0.6 mm, 0.7 mm, 0.8 mm, 0.9 mm, 1.0 mm, 1.1 mm, 1.2 mm, 1.3 mm, 1.4 mm, 1.5 mm, 1.6 mm, 1.7 mm, 1.8 mm, 1.9 mm, 2.0 mm, 2.1 mm, 2.2 mm, 2.3 mm, 2.4 mm, 2.5 mm, 2.6 mm, 2.7 mm, 2.8 mm, 2.9 mm, 3.0 mm, 3.1 mm, 3.2 mm, 3.3 mm, 3.4 mm, 3.5 mm, 3.6 mm, 3.7 mm, 3.8 mm, 3.9 mm, 4.0 mm, 4.5 mm, or 5.0 mm.

[0259] The device may have one or more fusion points. The points may be regularly spaced. The points may be regularly spaced to form a matrix array. The points may be irregularly or randomly spaced. In some embodiments, the points may be spaced at least 0.5 mm, 1.0 mm, 1.5 mm, 2.0 mm, 2.5 mm, 3.0 mm, 3.5 mm, 4.0 mm, 4.5 mm, 5.0 mm, 5.5 mm, 6.0 mm, 6.5 mm, 7.0 mm, 7.5 mm, 8.0 mm, 8.5 mm, 9.0 mm, 9.5 mm, or 10.0 mm center to center. In some embodiments, the points may be placed on top of each other and overlap.

[0260] The total surface area of ​​the dots may cover a portion of the surface area of ​​the membrane of the cell-retaining device. The surface area of ​​the membrane of the cell-retaining device covered by the dots may be a portion that does not interfere with its ability to maintain cell viability, cell function, and release of molecules from the interior of the device. In some embodiments, less than 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% of the membrane surface area may be covered by the dots.

[0261] The points can provide an adhesive constraint for limiting the bending of the film of the device away from each other. The point density and diameter can be selected to limit the deformation, bending or expansion of the film of the device away from each other during the filling of the device. The point diameter and density can affect the allowable filling volume by generating a series of opposites. For devices with the same size, a device with a shorter point spacing can have a higher point density than a device with a longer point spacing. Since the devices have the same point diameter and device size, a device with a shorter point spacing can have an internal volume that can be used for filling that is smaller than a device with a longer short spacing.

[0262] The size of the dots can be adjusted to control the volume within the cell containment device and the SA:V ratio. In some embodiments, the surface area of ​​the dots can be increased to reduce the volume within the cell containment device. Conversely, the increased surface area of ​​the channel can provide a higher SA:V ratio for the cell containment device. In some embodiments, the surface area of ​​the dots can be reduced to increase the volume within the cell containment device. The reduced surface area of ​​the dots can provide a lower SA:V ratio for the cell containment device.

[0263] The point can be produced according to various patterns. The point can be produced according to the pattern of the ability to fill the device uniformly and in the whole device. The point pattern can be designed to adjust the volume and / or amount of the cells that can be loaded into the device. The point pattern can be designed to limit the film expansion or bending of the device during filling the device with cells. In some embodiments, the device can have one point. In some embodiments, the device can have multiple points. In some embodiments, the device can have at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 points.

[0264] The thickness of the device during filling can also be controlled by applying an external constraint to the outer surface of the device. The external constraint can be placed on the outer surface of the membrane of the cell-retaining device and cover most or all of the surface of the membrane. The external constraint can physically limit the deformation of the membrane of the device away from each other during filling. The external constraint can be porous to help vent air displaced from the interior of the device as the device is filled. The external constraint can also be adjusted by using a spacer, wherein the spacer has a target distance between the porous constraints. The spacer can correspond to the total thickness of the device before filling.

[0265] The point can be produced by various methods to fuse the film in discrete points. The point can be produced by the fusion method using the spot welding process described herein. The point can be produced by placing adhesive on the desired position of the point on the first film and then contacting the first film with the second film. The bonding point can be built in multiple layers. In some embodiments, the bonding point is built in two layers. In some embodiments, adhesive can be placed on the film in an automatic or semi-automatic process. In some embodiments, an automatic dispenser can be programmed to dispense the adhesive of a specified volume or weight at a specified location. In some embodiments, the dispenser can be provided with information about the size of the film and the dot pattern to guide the adhesive to be dispensed on the film. The point can be produced by placing an adhesive in a combination with the spot welding process.

[0266] Many adhesives may be suitable for creating fusion points. The adhesive may be cyanoacrylate, polyurethane acrylate, UV curing epoxy, heat curing epoxy or a two part epoxy in which one element may be embedded in the film (monomer) and the other element may be applied in solution around it (crosslinker). Alternatively or in combination, a solvent may be used to partially dissolve the film junction to create the bond.

[0267] The properties of the adhesive and the pattern of adhesive placement can be used to adjust the dot diameter and density and thickness of the device during filling. The adhesive properties can affect the rate at which the adhesive invades the film. The first application can determine the effective diameter of the dot. The invasion rate can change with the viscosity of the adhesive. Depending on the chemical properties of the adhesive, the invasion rate may be affected by the charge and hydrophilization degree of the film. The coating, film and adhesive properties can adjust the density of the dot pattern together. Alternatively or additionally, the dot pattern can be controlled by a picosecond pulse adhesive dispenser, which adjusts these parameters according to time, and the dispensing performed by the dispenser can be nonlinear dispensing.

[0268] In some embodiments, the viscosity of the adhesive is about 200 cP to 450 cP. In some embodiments, the viscosity of the adhesive is at least 10 cP, 20 cP, 30 cP, 40 cP, 50 cP, 60 cP, 70 cP, 80 cP, 90 cP, 100 cP, 200 cP, 300 cP, 400 cP, 500 cP, 600 cP, 700 cP, 800 cP, 900 cP or 1000 cP.

[0269] Cell containment devices can be scaled to various sizes while maintaining various parameters constant. The device can be scaled to provide an equivalent microenvironment as in a device for human implantation, for example, in a device for mouse implantation. Various parameters important to the function of the device can be maintained constant. These parameters can be diffusion distances of relevant molecules (such as insulin), or SA:V ratios. Since the size of the device can affect the mechanical properties, the device can be designed to maintain the flexibility of the device without folding in the target implantation site. The enlarged device can be designed to be compatible with filling the device with a higher throughput because the filling process is also enlarged. Scaled device designs can be combined with updated fluid paths to improve the final seal of the device after filling. Scaled device designs can be combined with an integrated macro device framework for multiple devices, replacing e.g. Fig.74A and Fig.74B The basic flexible macro device framework shown in .

[0270] Provided herein is an ePTFE cell containment device comprising a first membrane and a second membrane opposite to and attached to the first membrane. The first membrane may include a first surface and a second surface. The first surface may include a plurality of channels and an opposite second surface having a surface area. The first membrane and the second membrane may form a closed compartment that provides a volume for containing cells within the ePTFE device.

[0271] The exemplary PVDF channel array device may attenuate under UV light.

[0272] Although the exemplary frameless ePTFE channel array device can achieve ePTFE transcription, in some cases, the device may be twisted in vivo. The exemplary frameless ePTFE channel array device adopts a stabilization frame and adopts straight and angled filling tubes respectively. The angled filling tube can provide improved fluid flow. In addition, the angled filling tube may increase the dead space in the filling tube attached to the film. Finally, the angled filling tube can reduce the necessity of laser vision during manufacturing. As seen, the angled filling tube is skewed relative to the symmetrical bisection plane of the film, while the straight filling tube is coplanar or parallel to the symmetrical bisection plane of the film.

[0273] Also provided herein is a method for manufacturing a cell containing device. The method may include providing a first membrane, forming a plurality of channels in the first membrane, and fusing a second membrane to the first membrane. The first membrane may include a first face and a second face. The second face may be opposite to the first face. The plurality of channels may be formed in the first face of the first membrane. The second membrane may be fused to the second face of the first membrane. The fusion of the second face of the first membrane with the second membrane may form a compartment. The compartment may be configured to contain cells between the second face of the first membrane and the second membrane.

[0274] The formation of the plurality of channels in the first surface of the first film may be performed by means of molds 1021, 1022. The molds may include Fig.86A The positive mold 1021 as seen in Fig.86B . The male mold 1021 can contact the second side of the first film, while the female mold 1022 can contact the first side of the film. Using the female mold 1022 can improve the 3D thermoforming of the film to produce a film with better thickness and porosity.

[0275] The upper portion of the exemplary channel array device can enable cells to be inserted into the device via an external filling tube. The lower portion of the exemplary channel array device includes the membrane. The external filling tube can include an angled filling tube as shown, wherein the filling tube is skewed relative to the symmetry bisecting plane of the membrane. The upper portion of the exemplary channel array device can also serve as a mechanical frame for support. The exemplary channel array device can have an internal volume of approximately 24 μl, a channel mass of approximately 50, a footprint of approximately 1x0.5 cm, a channel of 1 mm, and a channel-to-channel (CC) distance of approximately 1 mm. As described in accordance with Fig.87 As can be seen, the exemplary channel array device comprises a frame to which an insertion tube 1040 can be manually attached. Fig.87The exemplary channel array device shown in may include a rodent channel array device.

[0276] The dimple frame can make it possible to combine fluid paths within the channel array device. In addition, the dimple frame is easier to manufacture and does not require manual addition of insertion tubes. Multiple such channel array devices can be formed at one time. The multiple co-formed channel array devices can be easily filled, separated and sealed to allow autonomous and / or mechanically operated manufacturing. The dimple frame within the channel array device can be formed by selective laser sintering (SLS), injection molding, solvent casting, machining or any combination thereof. Such methods can allow a sufficiently high degree of control over the geometry and resolution of the dimple frame.

[0277] In addition, the dimple frame can be configured to impart torsional resistance, bending resistance, or both to the channel array device via its hoop strength. Such resistance can be provided by a dimple frame having a variable thickness, width, cross-sectional shape, or any combination thereof. In addition, the dimple frame sealed between the first membrane and the second membrane can be configured to be airtight to prevent contamination.

[0278] Finally, according to Fig.88A and Fig.88B , the dimple frame 1051 within the channel array device 1050 achieves a greater and more uniform mass flow rate than a channel array device 1050 with a filling port.

[0279] according to FIG. 89A to FIG. 89B , optimize the geometric design parameters of the channel array device 1070 to achieve increased effectiveness. The geometry of the channels in the channel array device 1070 can be defined by the cell chamber height (A), the fusion area (B), the opening diameter (C), the channel diameter (D), and the channel spacing (E).

[0280] The cell chamber height (A) can be measured as the maximum normal distance between the inner surface of the first membrane 1071 and the inner surface of the second membrane 1072. The cell chamber height (A) can be measured as the average of the maximum normal distance between the inner surface of the first membrane 1071 and the inner surface of the second membrane 1072 for all channels 1073 in the channel array device 1070. In some embodiments, the cell chamber height (A) is at least about 300 μm. According to Table 3 below, the cell chamber height (A) can be optimized to modify the diffusion flux, foreign body response, vascularization, non-invasive cell loading, and volume / footprint of the device.

[0281] The fusion area (B) can be calculated as the total surface area of ​​the fusion of the first film 1071 and the second film 1072. Alternatively, the fusion area (B) can be measured or correlated to the surface area of ​​the upper surface of the first film 1071 that is substantially parallel to the second film 1072. According to Table 3 below, the fusion area (B) can be optimized to modify the sealing integrity, foreign body reaction, and vascularization of the device.

[0282] In some embodiments, the device 1070 further includes an opening 1074 that penetrates the first membrane 1071 and the second membrane 1072 in the channel 1073. The opening 1074 can have an opening diameter (C). The opening diameter (C) can be measured according to the average, maximum, or minimum inner diameter of the opening 1074. The opening diameter (C) can be measured according to the average, maximum, or minimum opening diameter (C) of multiple openings 1074 in multiple channels 1073 in the device 1070. In some embodiments, the concentricity of the opening 1074 relative to the channel 1073 is at most 25% of the channel diameter (D). According to Table 3 below, the opening diameter (C) can be optimized to change the sealing integrity, foreign body reaction, and vascularization of the device.

[0283] In some embodiments, the channel diameter (D) is measured according to the maximum, minimum, or average inner diameter of the channel 1073. In some embodiments, the channel diameter (D) is measured according to the maximum, minimum, or average normal inner diameter of the channel 1073. In some embodiments, the channel diameter (D) is measured at the narrowest point in the channel. In some embodiments, the average diameter of the channel is about 400 μm to about 3,000 μm. According to Table 3 below, the channel diameter (D) can be optimized to modify the diffusion flux, foreign body response, vascularization, non-damaging cell loading, and volume / footprint of the device.

[0284] The channel spacing (E) can be measured according to the maximum, minimum or average distance between the inner surfaces of two adjacent channels 1073. The channel spacing (E) can be measured according to the maximum, minimum or average normal distance between the inner surfaces of two adjacent channels 1073. The channel spacing (E) can be measured according to the average of the maximum, minimum or average distance between the inner surfaces of two adjacent channels 1073 for each of the multiple channels 1073. In some embodiments, the center of each channel is separated from the center of another channel by a distance of about 75 μm to about 500 μm. According to Table 3 below, the channel spacing (E) can be optimized to change the diffusion flux, non-damage cell loading and volume / footprint of the device.

[0285] You can also follow Fig.89B The channel 1073 is characterized by its aspect ratio. The aspect ratio can be calculated as the ratio between the cell chamber height (A) and the channel diameter (D). The aspect ratio can be at least about 0.5.

[0286] [Table 3: Channel related parameters and functions]

[0287]

[0288]

[0289] according to Fig.89C , the geometry of the channel can be designed and optimized for use. The uniform cylindricality, circularity, and perpendicularity of the channel 1073 relative to the first surface of the first membrane 1073 enable optimal use and cell growth.

[0290] FIG. 90A to FIG. 90B The interior face of an exemplary first membrane showing an array of channels whereby the spacing and size of the channels achieve an increased surface to volume ratio, as described in Table 4 below. Fig.90A An exemplary channel array with a channel pitch of 150 μm is shown, and Fig.90B An exemplary channel array is shown with a channel pitch of 270 μm.

[0291] In some embodiments, at least one of the length and width of the channel array device 1070 is about 0.25 cm to about 3 cm. In some embodiments, each of the plurality of channels 1073 is substantially perpendicular to the first film. In some embodiments, the channels 1073 are arranged in a linear array. In some embodiments, the channels 1073 are arranged in an annular array. In some embodiments, the number of channels per unit area of ​​the device along the transverse plane is greater than about 50 / cm2. In some embodiments, the surface area to volume ratio of the device is at least about 40 cm-1. In some embodiments, the channel array device 1070 includes a compartment between the first film and the second film. The compartment may include a single continuous open space. The compartment may have a volume of about 8 μl to about 600 μl.

[0292] In some embodiments, the method of producing a channel array further comprises laser ablation of a portion of the first membrane and the second membrane within the plurality of channels. In some embodiments, the laser ablation removes a fused portion of the first membrane and the second membrane to form an opening.

[0293] In some embodiments, the concentricity of the opening relative to the channel is at most 25% of the diameter of the channel. Fig.91A (Left) Detailed image showing an exemplary laser-drilled channel array device whereby the openings have unacceptable concentricity greater than about 25% relative to the channels. Fig.91A (Right) Detailed image showing an exemplary laser drilled channel array device whereby the openings have an acceptable concentricity with respect to the channels of less than about 25%. Fig.91C , increased concentricity improves the uniformity of the width of the fusion zone around each opening, which achieves increased bond strength and seal. Additionally, increased concentricity increases the surface area to volume ratio and reduces the required size of the sealing area needed for strength and integrity, thereby reducing the sealing surface area, reducing the residual "shelf" that stimulates FBR and creates additional diffusion distance for concentrated blood vessels.

[0294] Openings and passages span in accordance with Fig.91B An exemplary array device of the alignment from left to right is shown in Fig.92 , whereby a concentricity value of 0 corresponds to perfect concentricity. For example, an opening formed using the laser ablation parameters associated with test 894 (center) has a much higher concentricity value than an opening formed using the ablation parameters associated with test 898 (right).

[0295] Fig.93A An image showing an exemplary laser drilled channel array device. Fig.93B Image showing the sealing interface of the channel array device. FIG. 94A to FIG. 94B High and low magnification images showing vascularization around an implanted channel array device. Fig.95 A diagram showing vascular host integration according to some embodiments. In some embodiments, at least one of the first membrane and the second membrane is configured to enable vascularization of cells within the device. In some embodiments, at least one of the first membrane and the second membrane is configured to enable vascularization of cells within the device in the absence of immunosuppressive therapy.

[0296] Equilibrium O of venous capillaries of exemplary channels at a pressure of 100 mmHg with channel diameters of 1,100 μm, 1,300 μm, and 1,500 μm and channel edge-to-edge distances of 150 μm and 250 μm. 2 Tension distribution is shown in Fig.96A In addition, the equilibrium O of the venous capillaries of the exemplary channel at a pressure of 45 mmHg was measured when the channel diameter was 1,100 μm, 1,300 μm, and 1,500 μm and the channel edge-to-edge distance was 150 μm and 250 μm. 2 Tension distribution is shown in Fig.96B middle.

[0297] In some embodiments, molding the plurality of channels with a mold includes thermoforming the plurality of channels. FIG. 97A to FIG. 97BA top perspective image and a cross-sectional image of an exemplary non-sintered film thermoformed at 360° C. in 7 minutes under a negative pressure of 3 psi are shown, respectively. As can be seen, the thermoforming process results in a formed channel without cracks or structural collapse. Such a thermoforming process achieves a significant curvature and a large cell chamber height 1150 of about 558 μm, which increases the surface area to volume ratio and provides sufficient space for accommodating cells. Although, thermoforming of the exemplary film does not result in bursting, according to Fig.98 , alternative membranes and processes can achieve greater fusion strength due to reduced sintering and greater forgeability. Such methods and membranes can also be capable of producing larger cell chamber heights 1150.

[0298] Fig.99A Arrays of channels with burst (left) and without burst (right) are shown. As seen above, thermoforming the exemplary film at 3.5 psi and 370° C. in 7 minutes yielded a flow rate of 100 Hz to 100 Hz. Fig.99B The channel with the largest cell chamber height is shown. Fig.100 and Table 6 below, Fig.99B The membrane achieves a good cell chamber height of about 672 μm and a cell volume of about 27 μl.

[0299] [Table 6: Membrane types and associated parameters]

[0300] Membrane Type Feature height (μm) Filling volume (μL) Theoretical cell dose A 450 ～16 1.34E+07 B 558 ～21 1.77E+07 C 672 ～27 2.27E+07

[0301] In addition, according to Fig.101 , thermoforming at a temperature of about 370°C achieves increased fusion peel resistance of the film, the channel of the film is used at a temperature of 474°C, a fusion time of 0.05 seconds and a Z deviation of 2.625 mm Fig.103 The fusion tool is used to form.

[0302] in addition, Fig.102A The results are shown in the case where the normalized enthalpy is 23.696 J / g and the start x is 321.25 °C and in the case where the normalized enthalpy is 23.141 J / g and the start x is 321.14 °C. Fig.99B Differential scanning calorimetry of an exemplary film of. In addition, Fig.102B Electron micrographs of the membrane are shown with (right) and without (left) sintering of the membrane. Sintering of the membrane can increase the stability of the cellular device.

[0303] Fig.103Illustrations and images showing exemplary hot fusion tools. The hot fusion tool may include a position-based fusion tool that is configurable to give a set number of fusion impacts, each impact lasting a set fusion time. In some embodiments, the set fusion time is less than about 1 second. In some embodiments, the fusion tool 1220 impacts each of one or more points on the first film 1 to 6 times. In some embodiments, the fusion tool 1220 impacts each of one or more points on the first film at most about 16 times. In addition, the temperature of the fusion head can be adjusted to change the properties of the channel and fusion formed thereby. In some embodiments, the set fusion temperature is about 250°C to about 600°C. In some embodiments, the fusion tool 1220 has an impact contact area of ​​at least about 0.07mm2. In some embodiments, the fusion tool 1220 contacts the side of the first film opposite to the thermoforming surface.

[0304] In some embodiments, the channel is formed by placing the first film and the second film in a frame and hitting one or more points on the first film with a fusion tool 1120. In some embodiments, the frame includes at least a portion of the outer edge of the first film and the second film. The first film and the second film can be generally parallel during fusion. The first film and the second film can be generally aligned, whereby the entire or most of the second film is covered by the first film. The first film and the second film can be separated by a gap distance. In some embodiments, the gap distance is about 300 μm to about 1,200 μm.

[0305] In some embodiments, the impact on the first film pierces the first film, the second film, or both, and fuses a portion of the first film to the second film. In some embodiments, at least one of the first film and the second film is substantially flat. In some embodiments, at least one of the first film and the second film comprises a non-sintered flat sheet.

[0306] It may be necessary to optimize the fusion impact parameters to prevent inconsistencies, such as Fig.104 Those shown in are not consistent, and some areas of the fusion are misaligned and inconsistent. Fig.105A The exemplary device shown in FIG. measures the Fig.105B Tensile peel strength tests suggest that fusion strength is inversely proportional to the amount of non-parallelism.

[0307] Although according to FIG. 106A to FIG. 106B Laser drilling openings in posterior fusion devices can improve the concentricity of the openings, but misalignment of such laser drilling tools may impart additional inconsistencies. Fig.97B, such errors can be corrected with adhesives, but the incorporation of load cells within the fusion probe and the use of robotic manufacturing and guidance equipment have improved the manufacturing method to achieve less rework time, scrap, and leakage. The incorporation of load cells allows for accurate calibration of the fusion force independent of tool geometry. The development of optimized, robot-enabled fusion parameters has increased processing speed and reduced heat shrinkage effects. Vision guidance systems have improved the concentricity of the fusion relative to the thermoforming channel.

[0308] An optional exemplary load sensitive thermal fusion tool may include a tip, a load cell, and a frame configured to hold a film. The thermal fusion tool may include a position based fusion tool that is configurable to impart a set fusion force within a set number of fusion strikes within a set fusion time. The load cell enables the fusion tool to fuse each point with the same force. Fig.107 An exemplary array device including a single row of channels formed by a thermal fusion tool is shown in FIG. Fig.108 , the shortest fusion time of about 0.05 seconds produced the strongest peel force fusion (.45N) between the films. Since the peel force indicates the fusion strength, a lower fusion time can increase the stability and service life of the device. Fig.109 , a complete device formed by a hot fusion tool at a fusion temperature of 800°F with one strike per location and a fusion force of about 6 pounds is shown in FIG. The circular uniform shape of the channels and Fig.110 The peel force stress-strain curve of the device shown in indicates that a high fusion strength of about 0.45N was confirmed.

[0309] exist Fig.111 An exemplary channel array device with channels made from 2, 4, 6, and 8 fusion hits. Fig.112 Although eight fusion hits produced the strongest fusion, four fusion hits produced the strongest fusion while leaving the channel intact.

[0310] Fig.113 A bar graph showing Fusion Strength / Peel Force (N) versus Fusion Force for an exemplary channel array device having a first deformed membrane and a second flat membrane with a fusion temperature of 800°F, a fusion time of 0.05 seconds, and a single fusion strike. While 6 and 12 lbs. fusion strike forces resulted in devices with higher peel forces of approximately 0.4 N and approximately 0.36 N, respectively, no observable relationship between the two variables was determined. The higher fusion strike forces may be less than ideal due to associated deterioration of the fusion tool tip and membrane frame. According to Fig.114, this correlation is confirmed, the figure shows that the exemplary array channel device including the membrane formed with a fusion force of 8 pounds exhibits membrane tearing, wrinkling, and oblong or double channels. Thus, a fusion force below 8 pounds is more ideal for the exemplary membrane.

[0311] Fig.115 The device where the channel was formed by 4 fusion strikes at a fusion force of 6 pounds was shown to be a stronger device (0.65 N) than the device where the channel was formed by 4 fusion strikes at a fusion force of 3 pounds (0.47 N), where both devices were formed at a fusion temperature of 800°F and a fusion time of 0.05 seconds. In addition, Fig.116 It was shown that devices formed from four hits at six pounds of force (~0.65) and from two hits at eight pounds of force (~0.7N) at a fusion temperature of 800°F and a fusion time of 0.05 seconds resulted in stronger devices than the exemplary devices formed from 1 or 2 hits at six pounds of force (~0.47N).

[0312] Fig.117A An image showing an exemplary array device having a 3x3 array of channels formed simultaneously, whereby each of the plurality of channels is struck once before a channel is struck a second time. Fig.117B , simultaneous impacts formed an exemplary device having a greater peel strength (0.9 N) than a device fused by sequentially impacting each channel twice before impacting another channel (0.75 N).

[0313] exist Fig.118 A comparison of the peel forces of exemplary fused films is shown in Table 7 below, where Film A was formed by striking the load cell fusion tool 2 times with a force of 8 pounds, Film B was formed by striking the load cell fusion tool 4 times with a force of 6 pounds, and Film C was formed using a non-load bearing unit fusion tool.

[0314] [Table 7: Comparison of fusion film peeling force]

[0315] membrane Fusion Impact Fusion (Ib) Highest average peel force (N) A 1 12 ~0.4N(single row) B 2 8 ~0.4~0.7N(single row) C 4 6 ~0.65N(single row)

[0316] exist Fig.119 Films A and C are shown in FIG, with Film A showing consistent uniform fusion. For Film A, no significant differences in the measured fusion strengths were found within the range of fusion forces applied. For Film B, fusion forces less than 8 pounds resulted in the strongest film with less tearing, wrinkling, and dragging. And for Film C, the highest fusion strength was measured at 6 pounds. Fig.112 Films A and C are compared in FIG, and film C clearly shows a more consistent and higher peel force.

[0317] Fig.121Low-resolution and high-resolution microscopy images of an exemplary film portion fused at a temperature of 800° F., within a fusion time of 0.05 seconds, with a fusion force of 6 pounds, and with four fusion strikes are shown. These images show that the exemplary film has a fusion strength of about 1.38 N, which is equivalent to the full strength of the unfused film.

[0318] Fig.122A Detailed image showing the fusion channel of the membrane with a seal size of approximately 170 + / - 8 um. Fig.122B The laser ablation and fusion areas show high concentricity. Fig.123 The fusion force / peel force (N) of exemplary pre-laser channel array devices fabricated using non-load cell enabled (Generation 1) and load cell enabled (Generation 2) fusion tools are shown, while the exemplary membrane formed from the membrane by the load cell enabled fusion tool exhibited a significantly higher (1.52 N) peel force.

[0319] Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present disclosure belongs.

[0320] As used herein, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Any reference to "or" herein is intended to include "and / or" unless otherwise specified.

[0321] As used herein, the term "about" refers to an amount that is approximately 10%, 5%, or 1% of the stated amount, including increments therein.

[0322] As used herein, the term "substantially perpendicular" refers to a relationship between two or more surfaces that are within 1 degree, 2 degrees, 3 degrees, 4 degrees, 5 degrees, 6 degrees, 7 degrees, 8 degrees, 9 degrees, 10 degrees, or increments therebetween of perpendicularity to each other.

[0323] As used herein, the term "substantially parallel" refers to a relationship between two or more surfaces that are within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 degrees, or increments therebetween of parallelism to each other.

[0324] As used herein, the phrases "at least one," "one or more," and "and / or" are open-ended expressions that are both conjunction and disjunction in operation. For example, each of the expressions "at least one of A, B, and C," "at least one of A, B, or C," "one or more of A, B, and C," "one or more of A, B, or C," and "A, B, and / or C" means A alone, B alone, C alone, A and B together, A and C together, B and C together, or A, B, and C together.

[0325] Although preferred embodiments of the present disclosure 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. Without departing from the present disclosure, those skilled in the art will now be able to imagine numerous variations, changes, and substitutions. It should be understood that various alternatives to the embodiments of the present disclosure described herein may be used to practice the present disclosure. It is intended that the following claims define the scope of the present disclosure and that the methods and structures within the scope of these claims and their equivalents are thereby encompassed.

[0326] [Example]

[0327] The present disclosure is further limited by the following non-limiting examples.

[0328] [Example 1: Hexagonal channel array device for evaluation in small animals]

[0329] This example describes the configuration of a hexagonal channel array device for evaluation in small animals. Figure 6 The display shows a presentation of hexagonal channel array devices with different sizes. The channel array device designed for testing in rats has an elongated hexagonal shape and measures 1.9 cm x 0.8 cm. The rat device has 93 channels and is 82 cm -1 The SA:V ratio is 97% safe volume and can hold a volume of 43 μl. The channel array device designed for testing in mice has a hexagonal shape and measures 0.64 cm x 0.55 cm. The mouse device has 19 channels and is 82 cm -1 The medium-sized device is 1.4 cm x 0.55 cm, has 49 channels, and is 83 cm -1 The SA:V ratio is 96%, the safe volume is 96%, and the volume of 24 μl can be maintained. The size of the device can be scaled while keeping the SA:V ratio constant.

[0330] [Example 2: Ultrastructural analysis of PVDF membrane after deformation step]

[0331] This embodiment describes the ultrastructural analysis of the PVDF membrane after the deformation step. The PVDF membrane undergoes the deformation step under a combination of pressure and temperature. The combination is 65psi and 135°C, 65psi and 150°C, 65psi and 165°C, 100psi and 135°C, 100psi and 150°C, 100psi and 165°C, 140psi and 135°C, 140psi and 150°C, and 140psi and 165°C. Fig.12 and Fig.16Scanning electron micrograph showing a cross section of a PVDF membrane after undergoing a deformation step. Fig.17 The measured channel depth or characteristic depth (μm) of a cell containment device is shown, wherein the deformation step is performed under different pressure and temperature conditions. The PVDF membrane has different channel depths depending on the temperature and pressure conditions, wherein a higher temperature of 165°C generally results in deeper channels. The size of the channel can be controlled by the temperature and pressure conditions. Control of the channel size is important because it determines the SA:V ratio of the cell containment device.

[0332] [Example 3: Analysis of PVDF cell containment device after fusion step]

[0333] This example describes the analysis of PVDF cell containment devices after the fusion step. The first PVDF membrane was subjected to a deformation step at 65 psi and 135°C, 65 psi and 150°C, 65 psi and 165°C, 100 psi and 135°C, 100 psi and 150°C, 100 psi and 165°C, 140 psi and 135°C, 140 psi and 150°C, or 140 psi and 165°C. Subsequently, the deformed membrane was fused to a second PVDF membrane at 225°C. Fig.13 Scanning electron micrographs showing cross-sections of PVDF cell containment devices after undergoing a fusion step. The first membrane deformed at 65 psi and 165° C., 100 psi and 150° C., 140 psi and 135° C., and 140 psi and 150° C. fuse to the second membrane. The first membrane deformed outside of those temperature and pressure parameters had poor fusion, with the seam between the first and second membranes clearly visible in the scanning electron micrographs.

[0334] Fig.19 Scanning electron micrographs showing cross-sections of cell containment devices with poor fusion, where the deformation steps were performed at various pressures and temperatures. Heat flow measurements by DSC of films deformed at 165°C and 140 psi showed a secondary peak at about 175°C, indicating crystalline rearrangement of this deformed film. Heat flow measurements of films deformed at 150°C and 100 psi and successfully fused to a second film did not show a secondary peak.

[0335] [Example 4: DSC analysis of fusion conditions and non-fusion conditions]

[0336] This example describes DSC analysis of fused and unfused conditions.The first PVDF membrane was subjected to a deformation step at 100 psi and 160°C or 100 psi and 173°C followed by quenching. Fig.15The heat flow measurements showing the film deformed at 100 psi and 160°C had a small shoulder and were similar to the baseline PVDF film, indicating that this film maintained a crystalline arrangement and would fuse to the second film. The heat flow measurements showing the film deformed at 100 psi and 173°C had a secondary peak, indicating that this film formed crystalline regions and would not fuse to the second film.

[0337] [Example 5: Surface profile of cell containment device]

[0338] This example describes the surface profile of a cell containment device. Fig.18 Surface interferometry of a cell containment device having a smooth surface with consistent geometry in its array of channels is demonstrated. The active zone has a smooth, porous surface, while the fusion zone has a flat surface.

[0339] [Example 6: Ultrastructural analysis of ePTFE after deformation step and fusion step]

[0340] This example describes the ultrastructural analysis of ePTFE after a deformation step and a fusion step.The ePTFE membrane was subjected to a deformation step at 30 psi and 340°C, 4 psi and 360°C, or 6 psi and 360°C. Fig.19 Scanning electron micrographs of ePTFE membranes after undergoing a deformation step are shown. The ePTFE membranes are capable of forming channels. In general, more deformation and fewer nodes are observed as temperature increases. Fig. 20 A fused ePTFE membrane was demonstrated that underwent a deformation step at 6 psi and 360°C and a fusion step within 5 minutes at 370°C. The ePTFE membrane requires a different temperature and pressure range for the deformation step and the fusion step compared to a PVDF membrane. The fabrication steps described herein can be applied to membranes of different materials to successfully form a cell containment device.

[0341] [Example 7: In vivo implantation of cell-containing devices in rats]

[0342] This example describes the in vivo implantation of cell containment devices in rats. The devices had similar channel diameters of about 350 μm, but different channel spacings. The low density device had a channel spacing between channels of about 500 μm, while the high density device had a channel spacing of about 200 μm. Fig.28 As shown in , cell containment devices with high and low channel density were implanted in vivo in various locations in normal rats via preperitoneal, intraomental, suprahepatic, and subcutaneous implantation. Fig.29Demonstrates vascularization around the cell-containing device after preperitoneal implantation. Microvessels were observed in each channel in the device. H&E histology of the vasculature indicated the presence of smooth muscle cells, which were found in the arteries. Fig.30 Demonstrating more vascularization and more branching of blood vessels as observed in high channel density devices compared to low channel density devices. Fig.31 The per-device counts of blood vessels and branches of low-channel density devices and high-channel density devices are shown. The high-channel density device has about 1000 blood vessels per device, as compared to about 500 blood vessels per device of the low-channel density device. The high-channel density device also has significantly more branches, about 500 branches per device, as compared to about 300 branches per device of the low-channel density device. The higher-channel density device causes more vascularization around the device. When more channels are added to the device, it may be necessary to balance the benefits of increased vascularization as seen in the higher-channel device with the integrity of the device. Channel density may affect the degree of vascularization around the cell-retaining device.

[0343] The cell containment device is filled with cells in preparation for in vivo implantation. Fig.42 H&E stained tissue sections showing various cell containment devices with continuous interior spaces completely filled with cells. A device made of 20 μm thick ePTFE membrane 5310 (top) is filled with cells 5320 in its single continuous interior space. This device can have a cell density of about 11 ng / cm per 10 minutes. 2 Modeled insulin diffusion rate. As shown at the bottom, a device made of 125 μm thick PVDF membrane 5330 is filled with cells 5320 in its single continuous interior space. This device can have a diffusion rate of about 6 ng / cm per 10 minutes. 2 Modeled insulin diffusion rate. Fig.43 An image showing an ePTFE cell containment device filled to maximum capacity with cells. On the top picture, an H&E stained tissue section shows a single continuous interior space of the device filled with cells throughout the device. In the lower left corner, a close-up image shows a microtissue containing cells in one recess of the interior space of about 150 μm in height and about 400 μm in width. A 5x magnified light-colored micrograph of an ePTFE cell containment device is shown in the lower right corner, demonstrating the continuous filling of its interior space in the manufactured cell containment device with channels. This demonstrates the ability to fill the entire interior space of a cell containment device with cells.

[0344] The cell containment device is implanted in vivo in a rat for an extended period of time, and the cells within the cell containment device survive during the extended in vivo implantation period. Fig.26 and Fig. 27Shown 20 days after implantation in the preperitoneal site in nude rats ( Fig.26 ) and 90 days ( Fig. 27 ) is a vasculature surrounding the cell containment device and in the channel. Fig.44 H&E stained tissue sections showing a PVDF cell containment device 5510 with cells 5520 after 90 days of in vivo implantation at the preperitoneal site in a rat. The SC islet cells 5520 within the device 5510 implanted in the preperitoneal site had high cellularity at 90 days and visible vascularization 5530 around the device and in the channels. Host tissue formed new tissue around the device and in the channels of the device. Fig.45 H&E stained tissue sections of PVDF cell containing devices 5610 filled with SC islet cells 5620 after 90 days of implantation in subcutaneous and preperitoneal sites in nude rats are shown, and the PVDF cell containing devices can react to foreign bodies. The SC islet cells 5620 implanted in the device 5610 in the subcutaneous and preperitoneal sites have high cell content at 90 days, indicating that the SC islet cells survive in the device at the two implantation sites for 90 days. Vascularization 5630 is observed around the device implanted at the subcutaneous and preperitoneal sites and in the channel of the device. Some of the vascularization is presented as having arteriolar-like features and is defined as a vascular wall with a thickness of multiple cell layers. The outer surface of the device is presented as being integrated into the host tissue at the two implantation sites, wherein there is a higher density of fibrosis around the device in the subcutaneous site but there is no significant effect on the survival of the cells in the device (top and lower left). The host tissue is also present in the channel of the device, providing support for vascular tissue and providing more mechanical stability for the device. This demonstrates the ability to implant a cell-containing device with cells in multiple implantation sites, maintain high cell viability of the cells within the device over extended periods of time, and have vascularization and new tissue formation around and within the device.

[0345] [Example 8: Cluster size affecting design]

[0346] This example describes how cluster size can affect the design.Cluster size refers to the size of the cell aggregates loaded into the device.

[0347] [Example 9: Deformation conditions of ePTFE membrane]

[0348] This example demonstrates the results of deformation conditions on an ePTFE membrane. Fig.41An ePTFE cell containment device formed by heat deformation at 360°C and 6 psi (formed; top) and subsequently hard cast with resin for cross-sectional imaging (bottom) is shown. The formed ePTFE membrane does not appear to retain its channel shape and has more fibril structure than nodes. The ePTFE membrane hard cast with resin appears to retain its channel shape after the deformation step.

[0349] [Example 10: T-peel test of two flat ePTFE membranes]

[0350] This example describes a T-peel test of sintered and non-sintered ePTFE membranes used to test the bond strength of fused ePTFE membranes. Fig.35 The breaking load (N) measured by the ASTM T-peel test is shown - according to the ASTM D882-08 standard for thin film tensile testing of two flat ePTFE membranes fused at different temperatures for different lengths of time (1 second, 5 seconds, 15 seconds). The ePTFE membranes were sintered (AS) or unsintered (AU) at 370°C for 7 minutes before fusion. As measured by DSC, the fusion temperature of the sintered ePTFE membrane is about 320°C to 325°C, and the melting temperature of the unsintered ePTFE membrane is about 340°C to 350°C. The two flat ePTFE membranes are fused together at various temperatures ranging from 302°C to 427°C within 1 second, 5 seconds or 15 seconds. After undergoing the ASTM T-peel test, the breaking or failure load (N) is recorded. In general, the fusion between sintered-sintered films (AS / AS) has the lowest failure load in the range of close to 0N to about 0.3N, as compared to non-sintered-non-sintered films (AU / AU) in the range of close to 0N to about 1N or non-sintered-sintered films (AU / AS) in the range of about 0.2N to 0.7N. In general, the fusion of AU / AU films has a higher failure load. In general, the failure load increases with increasing fusion temperature. In general, having at least one non-sintered film leads to a higher failure load.

[0351] [Example 11: T-peel test of ePTFE device]

[0352] This example describes the T-peel test of an ePTFE device. Fig.37A tool for ASTM T-peel testing according to the ASTM D882-08 standard for thin film tensile testing is shown. The tool was used to test fused cell-containing ePTFE devices and the tested devices at failure are shown. The figure shows the stress-strain curve of an ePTFE device with a sintered and deformed first membrane and a flat and non-sintered second membrane, the first membrane and the second membrane being fused in 0.05 seconds at 474°C. The failure strain of the ePTFE device was greater than 60% and a load of about 0.4N was achieved, which is about 78% of the load of a fused flat membrane with more fused area. This stress-strain curve demonstrates that even under high tensile strains, the fusion sites are fused and remain fused.

[0353] [Example 12: Burst pressure of ePTFE device]

[0354] This example describes measuring the burst pressure of a cell containment device. To test the strength of the seal of the device, the device was filled with water at 1 psi every 10 seconds and the failure or burst pressure was measured. Fig.39 A graph showing the failure filling pressure (psi) of a frameless ePTFE cell containment device and an ePTFE cell containment device (eCAD) prototype filled for burst pressure testing. For the PVDF prototype, the failure burst pressure was about 10 psi. For the ePTFE prototype, the failure burst pressure was about 11 psi. This burst pressure is much higher than the maximum load pressure of about 2 psi that the device may experience under typical filling conditions.

[0355] [Example 13: Cell containment device prototype]

[0356] This example describes a prototype of a cell containment device. Fig.14 Presentation of cell containment devices with scalloped perimeters and variations in channel dimensions to achieve various SA:V ratios. Fig.14 The cell containing device can be characterized by an overall height 1801 , an inner height 1802 , a membrane thickness 1803 , an inner spacing 1804 , an inner diameter 1805 , a through-hole inner diameter 1806 , and a through-hole spacing 1807 .

[0357] In some embodiments, the total height 1801 is about 400 μm to about 1,600 μm. In some embodiments, the total height 1801 is at least about 400 μm. In some embodiments, the total height 1801 is at most about 1,600 μm. In some embodiments, the total height 1801 is about 400 μm to about 600 μm, about 400 μm to about 850 μm, about 400 μm to about 1,000 μm, about 400 μm to about 1,200 μm, about 400 μm to about 1,400 μm, about 400 μm to about 1,600 μm, about 600 μm to about 850 μm, about 600 μm to about 1,000 μm, about 600 μm to about 1,200 μm, about 600 μm to about 1,400 μm, about 600 μm to about 1 In some embodiments, the total height 1801 is about 400 μm, about 600 μm, about 850 μm, about 1,000 μm, about 1,200 μm, about 1,400 μm, about 850 μm to about 1,600 μm, about 1,000 μm to about 1,200 μm, about 1,000 μm to about 1,400 μm, about 1,000 μm to about 1,600 μm, about 1,200 μm to about 1,400 μm, about 1,200 μm to about 1,600 μm, or about 1,400 μm to about 1,600 μm. In some embodiments, the total height 1801 is about 400 μm, about 600 μm, about 850 μm, about 1,000 μm, about 1,200 μm, about 1,400 μm, or about 1,600 μm. In some embodiments, inner height 1802 is about 300 μm to about 1,200 μm. In some embodiments, inner height 1802 is at least about 300 μm. In some embodiments, inner height 1802 is at most about 1,200 μm. In some embodiments, the inner height 1802 is about 300 μm to about 400 μm, about 300 μm to about 650 μm, about 300 μm to about 800 μm, about 300 μm to about 1,000 μm, about 300 μm to about 1,200 μm, about 400 μm to about 650 μm, about 400 μm to about 800 μm, about 400 μm to about 1,000 μm, about 400 μm to about 1,200 μm, about 650 μm to about 800 μm, about 650 μm to about 1,000 μm, about 650 μm to about 1,200 μm, about 800 μm to about 1,000 μm, about 800 μm to about 1,200 μm, or about 1,000 μm to about 1,200 μm. In some embodiments, inner height 1802 is 300 μm, about 400 μm, about 650 μm, about 800 μm, about 1,000 μm, or about 1,200 μm.

[0358] In some embodiments, the film thickness 1803 is about 50 μm to about 250 μm. In some embodiments, the film thickness 1803 is at least about 50 μm. In some embodiments, the film thickness 1803 is at most about 250 μm. In some embodiments, the film thickness 1803 is about 50 μm to about 75 μm, about 50 μm to about 100 μm, about 50 μm to about 125 μm, about 50 μm to about 150 μm, about 50 μm to about 175 μm, about 50 μm to about 200 μm, about 75 μm to about 100 μm, about 75 μm to about 125 μm, about 75 μm to about 150 μm, about 75 μm to about 175 μm, about 75 μm to about In some embodiments, the film thickness 1803 is about 50 μm, about 75 μm, about 100 μm, about 125 μm, about 150 μm, about 175 μm, about 200 μm, about 175 μm, about 200 μm, about 150 μm, about 175 μm, about 200 μm, about 175 μm, about 200 μm, about 150 μm to about 200 μm, or about 200 μm. In some embodiments, the film thickness 1803 is about 50 μm, about 75 μm, about 100 μm, about 125 μm, about 150 μm, about 175 μm, about 200 μm, or about 250 μm.

[0359] In some embodiments, the inner spacing 1804 is about 40 μm to about 500 μm. In some embodiments, the inner spacing 1804 is at least about 40 μm. In some embodiments, the inner spacing 1804 is at most about 500 μm. In some embodiments, the inner spacing 1804 is about 40 μm to about 60 μm, about 40 μm to about 80 μm, about 40 μm to about 100 μm, about 40 μm to about 150 μm, about 40 μm to about 200 μm, about 40 μm to about 270 μm, about 40 μm to about 350 μm, about 40 μm to about 400 μm, about 40 μm to about 500 μm, about 60 μm to about 80 μm, about 60 μm to about 100 μm, about 60 μm to about 150 μm, about 60 μm to about 200 μm, about 60 μm to about 270 μm, about 60 μm to about 350 μm, about 60 μm to about 400 μm, about 60 μm to about 500 μm, about 80 μm to about 100 μm, about 80 μm to about 150 μm, about 80 μm to about 200 μm, about 80 μm to about 270 μm, about 80 μm to about 350 μm, about 80 μm to about 400 μm, μm, about 80 μm to about 500 μm, about 100 μm to about 150 μm, about 100 μm to about 200 μm, about 100 μm to about 270 μm, about 100 μm to about 350 μm, about 100 μm to about 400 μm, about 100 μm to about 500 μm, about 150 μm to about 200 μm, about 150 μm to about 270 μm, about 150 μm to about 350 μm, about 150 μm to about 400 μm , about 150 μm to about 500 μm, about 200 μm to about 270 μm, about 200 μm to about 350 μm, about 200 μm to about 400 μm, about 200 μm to about 500 μm, about 270 μm to about 350 μm, about 270 μm to about 400 μm, about 270 μm to about 500 μm, about 350 μm to about 400 μm, about 350 μm to about 500 μm, or about 400 μm to about 500 μm. In some embodiments, the inner spacing 1804 is about 40 μm, about 60 μm, about 80 μm, about 100 μm, about 150 μm, about 200 μm, about 270 μm, about 350 μm, about 400 μm, or about 500 μm.

[0360] In some embodiments, inner diameter 1805 is about 300 μm to about 1,600 μm. In some embodiments, inner diameter 1805 is at least about 300 μm. In some embodiments, inner diameter 1805 is at most about 1,600 μm. In some embodiments, inner diameter 1805 is about 300 μm to about 500 μm, about 300 μm to about 700 μm, about 300 μm to about 900 μm, about 300 μm to about 1,100 μm, about 300 μm to about 1,300 μm, about 300 μm to about 1,600 μm, about 500 μm to about 700 μm, about 500 μm to about 900 μm, about 500 μm to about 1,100 μm, about 500 μm to about 1,300 μm, about 500 μm to about In some embodiments, inner diameter 1805 is about 300 μm, about 500 μm, about 700 μm, about 900 μm, about 1,100 μm, about 1,300 μm, about 700 μm to about 1,600 μm, about 900 μm to about 1,100 μm, about 900 μm to about 1,300 μm, about 900 μm to about 1,600 μm, about 1,100 μm to about 1,300 μm, about 1,100 μm to about 1,600 μm, or about 1,300 μm to about 1,600 μm. In some embodiments, inner diameter 1805 is about 300 μm, about 500 μm, about 700 μm, about 900 μm, about 1,100 μm, about 1,300 μm, or about 1,600 μm.

[0361] In some embodiments, the through-hole inner diameter 1806 is about 100 μm to about 600 μm. In some embodiments, the through-hole inner diameter 1806 is at least about 100 μm. In some embodiments, the through-hole inner diameter 1806 is at most about 600 μm. In some embodiments, the through-hole inner diameter 1806 is about 100 μm to about 200 μm, about 100 μm to about 300 μm, about 100 μm to about 400 μm, about 100 μm to about 500 μm, about 100 μm to about 600 μm, about 200 μm to about 300 μm, about 200 μm to about 400 μm, about 200 μm to about 500 μm, about 200 μm to about 600 μm, about 300 μm to about 400 μm, about 300 μm to about 500 μm, about 300 μm to about 600 μm, about 400 μm to about 500 μm, about 400 μm to about 600 μm, or about 500 μm to about 600 μm. In some embodiments, the through-hole inner diameter 1806 is about 100 μm, about 200 μm, about 300 μm, about 400 μm, about 500 μm, or about 600 μm.

[0362] In some embodiments, the through-hole spacing 1807 is about 100 μm to about 400 μm. In some embodiments, the through-hole spacing 1807 is at least about 100 μm. In some embodiments, the through-hole spacing 1807 is at most about 400 μm. In some embodiments, the through-hole spacing 1807 is about 100 μm to about 200 μm, about 100 μm to about 300 μm, about 100 μm to about 400 μm, about 200 μm to about 300 μm, about 200 μm to about 400 μm, or about 300 μm to about 400 μm. In some embodiments, the through-hole spacing 1807 is about 100 μm, about 200 μm, about 300 μm, or about 400 μm.

[0363] In one embodiment, the device may have a channel 1810 with an inner diameter 1805 of 800 μm and an inner height 1802 of 650 μm, and a through hole 1820 with a through hole inner diameter 1806 of 300 μm. This device may have an overall height 1801 of 850 μm, a membrane thickness 1803 of 100 μm, and an internal spacing 1804 between channels 1810 of 270 μm. This device may have a 77 cm -1 In another embodiment, the device may have a channel 1810 with an inner diameter 1805 of 650 μm and an inner height 1802 of 650 μm, and a through hole 1820 with an inner diameter of 400 μm. This device may have an overall height 1801 of 850 μm, a membrane thickness 1803 of 100 μm, and an internal spacing 1804 between channels 1810 of 80 μm. This device may have a 138 cm -1 The size and spacing of the channels 1810 can be adjusted to achieve different SA:V ratios. The device can have a through-hole spacing 1807 of about 200 μm.

[0364] [Example 14: Human device prototype]

[0365] This example describes a prototype of a cell containment device assembled onto a frame for human use. Fig.40 A prototype of three cell containment devices assembled to a frame for human use is shown. The cell containment devices may be hexagonal in shape. The frame may have dimensions of approximately 26.9 mm x 75.6 mm and may be oval in shape with holes in the spaces between the cell containment devices. The cell containment devices may be approximately 850 μm thick with a 20 cm 2 The assembled device can hold about 400 million cells with a cell mass volume of about 350 μL. These cells can be capable of producing insulin.

[0366] [Example 15: Surface modification of membrane]

[0367] This embodiment describes a method for modifying the surface of a membrane. The membrane can be treated to impart hydrophilic properties on a membrane having a hydrophobic surface. The surface of the membrane can be modified by cross-linking a polymer having a hydrophilic property. The polymer can also be biocompatible and form a biocompatible coating on the membrane. In one embodiment, hydroxypropyl acrylate (HPA) and tetraethylene glycol diacrylate (TEGDA) can be cross-linked to the membrane with ammonium persulfate (APS) initiator during thermal polymerization. Fig.41 An example of a protocol for forming a hydrophilic coating on the surface of a membrane is shown. The ePTFE device is soaked in 100% ethanol and then soaked in 30% ethanol for 3 minutes. The device is soaked in 3% HPA, 2% TEGDA, 1% APS in 30% ethanol for 5 minutes and heated from room temperature to 80°C. Subsequently, the device is boiled in 100% ethanol for 5 minutes, soaked in Milli-Q water or ultrapure water for 30 minutes, and dried. Fig.38 An ePTFE cell containment device is shown after hydrophilic coating treatment immersed in water. Fig.41 Electron micrograph showing an ePTFE device after surface modification.

[0368] [Example 16: Dot diameter and density of the device]

[0369] This embodiment describes the point diameter and density (e.g., center to center spacing) of the device. The bonding points are built in 2 layers. The device is turned over by hand to produce a pattern of bonding points for limiting the thickness of the device during cell filling. A pattern of 37 points is deposited in 2 continuous layers, each point diameter is about 1.25mm, and the center to center spacing is 3.3mm. First, the pattern is formed on the film by applying adhesive in 0.05 seconds of application and curing each point individually for 1 second. A smaller second layer is applied on the top of the first pattern with 0.03 seconds of dispensing time, and the second layer is made uncured. The peripheral adhesive is then placed as described herein, and the film is placed on the top of the uncured adhesive by machine vision. After 1 second of intrusion time, the entire assembly is cured by first curing the periphery and then sweeping the internal area to cure the point pattern for 28 seconds and 112 seconds respectively. The completed device is then manually removed from the assembly platform and placed in a secondary container to perform post-assembly thermal curing at 37°C within 2 hours.

[0370] [Example 17: Point diameter of device and filling parameters]

[0371] This embodiment describes the filling volume of the device with various point configurations. Point diameter and density (e.g., center to center spacing) can affect the permissible filling volume by producing a series of columns. As described in Example 16, the device is prepared to have various point configurations. The device is prepared to have a certain point pattern, each point diameter is about 1.25mm and the point spacing is 2.6mm, 3.3mm or 4.4mm, wherein the center of one point is separated from the center of its adjacent point by the point spacing distance to place. For devices of the same size, the device with a shorter point spacing may have a point density higher than the device with a longer point spacing. Since the device has the same point diameter and device size, the device with a shorter point spacing can have an internal volume that can be used for filling that is smaller than the device with a longer point spacing. Subsequently, when there is no constraint or an external constraint outside the device, the cell suspension is loaded to the device. The external constraint can prevent the film of the device from expanding outward away from each other. Figure 2 The amount of cells loaded into the device is shown to vary with the spot spacing and the presence of constraints. When the spot spacing is reduced, fewer cells are loaded. The cell amount is 108x10 in a device with a spot spacing of 4.4 mm and loaded without constraints. 6 The number of cells is reduced to 42x10 in a device with a 2.6 mm point pitch and loaded with external constraints. 6 For devices with the same inter-point spacing, the larger number of cells loaded into the device is reduced when loaded with external constraints.

[0372] [Example 18: Adhesion points on cell containment device]

[0373] In the process of producing points on the cell-containing device, the device can be turned over by hand to produce a pattern of adhesive points for limiting the thickness of the device during cell filling. For example, a pattern of 37 points is deposited in 2 consecutive layers, each point diameter is about 1.25mm, and the center-to-center spacing is 3.3mm. First, the pattern is formed on the film by applying the adhesive in 0.05 seconds of application and curing each point individually for 1 second. A smaller second layer is applied on the top of the first pattern with a 0.03 second dispensing time, and the second layer is uncured. The peripheral adhesive is then placed on the film, and the film is placed on top of the uncured adhesive by machine vision. After a 1 second intrusion time, the entire assembly is cured by first curing the periphery and then sweeping the internal area to cure the dot pattern for 28 seconds and 112 seconds respectively. The completed device is then manually removed from the assembly platform and placed in a secondary container for post-assembly heat curing at 37°C within 2 hours.

[0374] [Example 19: In vivo implantation of ultrathin device]

[0375] This example describes the implantation of an ultrathin cell containment device into the NOD scid gamma (NSG) mouse model (an immunodeficient mouse model). Diabetes was induced in NGS mice, where Figures 61A to 61C As shown in , the blood glucose level of mice increased to above 400 mg / dL. After diabetes induction, the ultrathin device was implanted in the epididymal fat pad of mice. Fig.61A Examples of implanted ultrathin devices are shown. These implanted ultrathin devices include a high-throughput ePTFE membrane with a hydrophilic coating and 8 million SC pancreatic islet cells. After implantation of the ultrathin device, Fig.61A As shown in , blood glucose levels in all tested mice decreased to about 100 mg / dL and approached the level before diabetes induction within 90 days until the ultrathin device was removed or explanted. The explanted ultrathin device was analyzed by histology. Fig.61B and Fig.61C Demonstrated high cell density within the device, including the core of the device, 90 days after implantation in the NSG mouse model.Ultrathin devices populated with islet cells can be implanted in diabetic subjects and reduce and normalize blood glucose levels in the subject over an extended period of time.

[0376] [Example 20: In vivo implantation of AS-1 ultrathin device]

[0377] This example describes the condition of cells within the ultrathin device 30 days and 3 months after implantation in a mouse model in vivo. Fig.62 Low magnification image showing a histological section of a cell-filled ultrathin device 30 days after in vivo implantation in a mouse model. The ultrathin device was made of AS-1 membrane and filled with SC-islet cells called SEM-01. Fig.63 High magnification images of the tissue sections are shown. After 30 days in vivo the cells were evenly distributed and viable in the device.

[0378] [Example 21: In vivo implantation of endocrine cells in an ultrathin device]

[0379] This example describes the cellular prototype of endocrine cells in an ultrathin device 3 months after implantation in a mouse model in vivo. Fig.64 Microclusters of endocrine cells are shown prior to encapsulation and filling into cell containment devices. Fig.65 Shows stained histological images of ultrathin devices populated with micromasses of endocrine cells 3 months after in vivo implantation in mice. Blue staining indicates cell nuclei, orange-brown staining indicates the presence of C-peptide, and pink staining indicates the presence of glucagon. Fig.64Orange-brown and red staining in the micromasses in the Figure 2 indicates the presence of active endocrine cells secreting C-peptide and glucagon. Fig.65 The orange-brown and pink staining in the micromasses in indicated that the endocrine cells in the ultrathin device remained viable and active and maintained their endocrine prototype, secreting C-peptide and glucagon during 3 months of implantation in vivo.

[0380] [Example 22: Intraperitoneal glucose tolerance test using an ultrathin device]

[0381] This example describes an intraperitoneal glucose tolerance test performed using various configurations of the ultrathin device. Fig.66 The level of serum C-peptide and total insulin content in mice implanted with endocrine cells or ultrathin devices filled with endocrine cells are shown. The test groups are D601mcRA2 endocrine cells implanted under the membrane (group A), ultrathin devices with AS-1 membranes and filled with D601 cells (group B), and ultrathin devices with Class A ePTFE membranes and filled with D601 cells (group C). The cells or devices are implanted in mice and undergo an intraperitoneal glucose tolerance test. Serum C-peptide levels at baseline (blue) and under 30-minute glucose stimulation (orange) and total insulin content in the transplants are measured. All groups show an increase in serum C-peptide levels under glucose stimulation, indicating insulin production stimulated by glucose. Group A has a serum C-peptide level of about 100pM at baseline and a serum C-peptide level of about 250pM under glucose stimulation, and has a total insulin content of about 700μg. Group B had a serum C-peptide level of about 100 pM at baseline and a serum C-peptide level of about 200 pM under glucose stimulation, and had a total insulin content of about 500 μg. Group C had a serum C-peptide level of about 50 pM at baseline and a serum C-peptide level of about 600 pM under glucose stimulation, and had a total insulin content of about 550 μg. This example shows evidence of insulin control using an increase in insulin production as measured by increased serum C-peptide levels.

[0382] [Example 23: Host-ultrathin device interaction in a nude mouse model]

[0383] This example describes host-ultrathin device interactions following device implantation in a nude mouse model. Fig.67H&E staining images of explants of ultrathin devices comprising a coated selectively permeable membrane and filled with SC-islet cells after 3 months at the preperitoneal site in nude mice are shown. The images show a reduction in foreign body response (FBR) due to the lack of macrophages and macrophage fusion and angiogenesis in the ultrathin device with a selectively permeable membrane having a hydrophilic coating. The images also show the separation of host tissue from the interior of the ultrathin device and the concentration of viable cells within the device.

[0384] [Example 24: Cell viability and prototype in implanted devices populated with SC-islet cells]

[0385] This example describes cell viability and maintenance of the prototype after 12 weeks of implantation of ultrathin devices populated with SC-islet cells in a nude mouse model. Fig.68 H&E stained images of an ultrathin device filled with 16 million SC-islet cells 12 weeks after implantation at the preperitoneal site in a nude mouse model are shown. The images show high levels of cell viability in the device core after 12 weeks in vivo. Fig.69 Serum C-peptide levels in 4 different mice implanted with ultrathin devices filled with SC-islet cells over 60 minutes after glucose was administered to the peritoneal cavity are shown. Serum C-peptide increased over time from about 30 pM to 100 pM at 0 minutes to about 1000 pM or more in the four mice. This indicates that the SC-islet cells implanted in the ultrathin devices in the mice are active and maintain their ability to produce insulin.

[0386] [Example 25: Biocompatibility of Ultrathin Devices Implanted in an Immunocompetent Mouse Model]

[0387] This example describes the biocompatibility of an ultrathin device with an AS-1 membrane implanted in an immunocompetent black 6 mouse model. Empty mouse-sized ultrathin devices coated with an AS-1 membrane were placed subcutaneously in black 6 mice to assess the baseline host response to the materials of the ultrathin device. After 1 month, the device was assessed for maintenance of device integrity in terms of host tissues and cells and for foreign body response (FBR). Fig.70A and Fig.70B The lack of cells inside the device and the lack of FBR are shown. FBR has been described to peak within the first month (Beets, 1998). The images indicate that the integrity of the device is maintained in vivo and that the device materials and coatings are biocompatible and do not cause FBR.

[0388] [Example 26: Implantation of ultrathin devices in diabetic NSG mouse model]

[0389] This example describes the implantation of ultrathin devices with AS-1 membrane and encapsulated rat islet cells in a diabetic NSG mouse model.Ultrathin devices with AS-1 membrane and filled with 400 IEQ rat islet cells were implanted in diabetic NSG mice for 90 days. Fig.71 H&E stained images showing ultrathin devices with viable, intact rat islet cells after 90 days in vivo. Fig.72 Blood glucose levels before and after 90 days of ultra-thin device implantation are shown. After inducing diabetes, animals were administered insulin pellets to control blood glucose within the first 10 to 20 days of the experiment. After implanting the ultra-thin device (labeled as "implant"), blood glucose levels were reduced from more than 400 mg / dL to between about 100 mg / dL and about 300 mg / dL after 90 days of implantation. After the device was removed from mice, blood glucose levels increased. This indicates that the implanted ultra-thin device can provide extended glucose control.

[0390] [Example 27: Cell filling of ultrathin devices]

[0391] Fig.73 and FIG. 74A to FIG. 74B Various configurations of ultrathin devices are shown. An ultrathin device designed for implantation in a mouse model can be filled with 8 million cells and have a fusion point in the center of the device. An ultrathin device designed for human implantation can have no fusion point or point array and be filled with 133 million cells.

[0392] [Example 28: Mass flow rate of ultrathin device with dots]

[0393] Fig.76A An ultrathin device for human implantation is presented having an array of spot welds that provide adhesive constraints for preventing the two membranes of the device from bending away from each other when the device is filled. Fig.76B An arrangement is shown with a porous metal platen to provide an external constraint to an ultra-thin device to reduce bending of the two membranes of the device away from each other when the device is filled. The external porous constraint can be used to fill any cell-retaining device, including ultra-thin devices and ultra-thin devices with spot welds. Fig.77 Mass flow rates measured from filling of an ultrathin device with 3.3 mm point spacing for human implantation with and without external porous constraints are shown. 3 The mass flow rate measured in units of 100 sccm / min or standard milliliters per minute (sccm) is similar with or without an external porous restriction, reaching a peak of about 7.5 or 8 sccm at about 10 seconds and decreasing over time. After reaching the initial peak, the mass flow rate is slightly higher without an external porous restriction. Fig.78 Display and Fig.76AH&E staining images of cell distribution throughout an ultrathin device with adhesive constraints similar to the device shown in . This demonstrates the ability to uniformly populate a device with adhesive constraints throughout the device.

[0394] [Example 29: Cell filling of ultrathin devices without or with dots]

[0395] This example describes cell filling of ultrathin devices without or with dots. Fig.79A and Fig.79B Two configurations of a hexagonal ultrathin device are shown with no dots (A) and with a 3.3 mm dot array matrix (B) within the device. Fig.80 Display can fill in the area without points (such as Fig.79A ) and a 3.3 mm dot array matrix (such as Fig.79B The amount of cells in a single ultrathin device (in ). A device without dots can be filled with about 120 million cells, and a device with a 3.3 mm dot matrix can be filled with about 80 million cells. This demonstrates that the dot array matrix can be used to adjust cell loading and constrain membrane expansion or bending of the device during filling of the device with cells.

[0396] [Example 30: Cell filling of ultrathin devices with constraints]

[0397] This example describes cell filling of ultrathin devices with and without porous confinement. Fig.81A and Fig.81B Two configurations of a hexagonal ultrathin device with a 3.3 mm dot array matrix are shown filled without any constraints (A) and with a porous press plate spaced apart by 400 μm spacers (B). Fig.82 Display without any constraints (such as in Fig.81A in) and in cases with porous constraints (such as in Fig.81B The amount of cells that can fill a single ultrathin device in the case of (). The device without any constraints can be filled with about 87 million cells, and the device with porous constraints can be filled with about 80 million cells. This demonstrates that porous constraints can reduce membrane expansion and bending during filling of the device. The constraints can also be adjusted by using spacers with a target distance between porous constraints.

[0398] [Example 31: In vivo implantation in a minipig model]

[0399] This example describes minipig implantation studies of human-sized ultrathin devices. Ten minipigs were implanted with empty ultrathin devices, or ultrathin devices filled with SC-islet cells (SEM-01) or porcine islets at preperitoneal or subcutaneous sites. Fig.83AAn example of an ultra-thin device with a 2.6 mm dot pitch, using a human single module design, implanted in a miniature pig is shown. Fig.83B The target preperitoneal or subcutaneous implantation site in miniature pigs is demonstrated approximately 3 inches distal to the midline and avoiding the rib margin. Fig.83C Demonstrates subcutaneous dissection using a Bovie electrocautery in preparation for device implantation. Fig.83D Demonstration of preperitoneal dissection performed with a lighted retractor in preparation for device implantation. The dissection of the implantation site can be tailored for different implantation methods. Fig.84A Example showing subcutaneous implantation of an ultrathin device. Fig.84B Example demonstrating preperitoneal placement of an ultrathin device. Fig.85 Examples of minipigs 2 weeks after subcutaneous (SQ) and preperitoneal (PP) implantation of the ultrathin device are shown. The images show no clear evidence of inflammation around the implantation site, and the animals exhibited no signs of distress or pain.

[0400] This manual also includes the following contents:

[0401] 1. A cell containing device, comprising:

[0402] (a) a first membrane having a first surface including a plurality of channels and a plurality of second surfaces opposite the first surface; and

[0403] (b) a second film, the second film being opposite to the first film and attached to the plurality of the second surfaces of the first film;

[0404] wherein the first membrane and the second membrane form a closed compartment having a surface area to volume ratio of at least about 40 cm -1 ,and

[0405] Wherein the enclosed compartment provides a volume for containing cells within the device.

[0406] 2. The device of embodiment 1, wherein the compartment comprises a single continuous open space.

[0407] 3. The device of embodiment 1, wherein the volume is about 8 μl to about 1,000 μl.

[0408] 4. The device of embodiment 1, wherein at least one of the length and width of the device is from about 0.25 cm to about 3 cm.

[0409] 5. The device of embodiment 1, wherein the thickness of the device is at least about 300 μm.

[0410] 6. The device of embodiment 1, wherein the plurality of channels are substantially perpendicular to the first membrane.

[0411] 7. The device of embodiment 1, wherein the plurality of channels are arranged in a linear array.

[0412] 8. The device of embodiment 1, wherein the plurality of channels are arranged in a circular array.

[0413] 9. The device of embodiment 1, wherein the average diameter of the plurality of channels is about 400 μm to about 3,000 μm.

[0414] 10. The device of embodiment 9, wherein the diameter is measured at the narrowest point in the plurality of channels.

[0415] 11. The device of embodiment 1, wherein the center of each of the plurality of channels is separated from the center of another channel by a distance of about 75 μm to about 500 μm.

[0416] 12. The device of embodiment 1, wherein the channel has a height to diameter ratio of at least about 0.2.

[0417] 13. The device of embodiment 1, wherein the number of channels per unit area of ​​the device along the transverse plane is greater than about 50 / cm 2 .

[0418] 14. The device of embodiment 1, wherein at least one of the first membrane and the second membrane comprises a plurality of nodes interconnected by a plurality of fibrils.

[0419] 15. The device of embodiment 1, wherein at least one of the first membrane and the second membrane comprises PVDF, PTFE, ePTFE, PCL, PE / PES, PP, PS, PMMA, PLGA, PLLA, or any combination thereof.

[0420] 16. The device of embodiment 1, further comprising an opening in the channel extending through the first membrane and the second membrane.

[0421] 17. The device of embodiment 16, wherein the concentricity of the opening relative to the channel is at most 25% of the diameter of the channel.

[0422] 18. The device of embodiment 1 further comprises a frame configured to house the device.

[0423] 19. The device of embodiment 18, wherein the frame is configured to house a plurality of cell-retaining devices.

[0424] 20. The device of embodiment 18, wherein the frame comprises a flexure mechanism configured to prevent buckling of the cell containment device.

[0425] 21. The device of embodiment 1 further comprises a cell population.

[0426] 22. The device of embodiment 21, wherein the cell population is an insulin-secreting population.

[0427] 23. The device of embodiment 21, wherein the cell population is stem cell-derived cells capable of achieving glucose-stimulated insulin secretion (GSIS).

[0428] 24. The device of embodiment 1, further comprising a coating comprising a hydrophilic polymer.

[0429] 25. The device of embodiment 1, wherein the insulin diffusion coefficient of the device is about 2×10-6 cm 2 / s to about 1×10-5 cm 2 / s.

[0430] 26. The device of embodiment 1, wherein the maximum oxygen diffusion distance of the device is less than about 150 μm.

[0431] 27. The device of embodiment 1, wherein the first film and the second film are fused by a fusion peel force of at least about 0.4N.

[0432] 28. The device of embodiment 1, wherein at least one of the first membrane and the second membrane is semi-permeable.

[0433] 29. The device of embodiment 28, wherein the semi-permeability of the first membrane, the second membrane, or both is configured to protect the cells from immune attack.

[0434] 30. The device of embodiment 29, wherein the semi-permeability of the first membrane, the second membrane, or both is configured to protect the cells from immune attack in the absence of immunosuppressive therapy.

[0435] 31. The device of embodiment 1, wherein at least one of the first membrane and the second membrane is configured to achieve vascularization of the cells within the device.

[0436] 32. The composition of embodiment 31, wherein at least one of the first membrane and the second membrane is configured to achieve vascularization of the cells within the device in the absence of immunosuppressive therapy.

[0437] 33. A cell containing device, comprising:

[0438] (a) a first membrane having a first surface including a plurality of channels and a plurality of second surfaces opposite the first surface; and

[0439] (b) a second film, the second film being opposite to the first film and attached to the plurality of the second surfaces of the first film;

[0440] wherein the first membrane and the second membrane form a closed compartment

[0441] wherein the enclosed compartment provides a volume for housing between 1 million and 1 billion insulin-producing cells within the device, and wherein the membrane permits diffusion of insulin from the device while retaining the insulin-producing cells within the device.

[0442] 34. A composition comprising insulin-producing cells and a device housing the insulin-producing cells, wherein the device, after implantation in an individual, releases insulin while retaining the insulin-producing cells in the device and promotes vascularization of tissue in and around the device.

[0443] 35. The composition of embodiment 34, wherein no immunosuppressant is administered to the individual during implantation or vascularization of the device.

[0444] 36. The composition of embodiment 34, comprising 1 million to 1 billion insulin-producing cells.

[0445] 37. The composition of embodiment 34, wherein the thickness of the device is at least about 300 um.

[0446] 38. A composition as described in embodiment 34, wherein the device comprises a membrane comprising a plurality of nodes interconnected by a plurality of fibrils.

[0447] 39. A method for manufacturing a cell containment device, the method comprising:

[0448] (a) providing a first film having a first side and an opposite second side;

[0449] (b) forming a plurality of channels in the first face of the first membrane; and

[0450] (c) fusing a second membrane to the second side of the first membrane to form a compartment for accommodating cells between the second side of the first membrane and the second membrane.

[0451] 40. The method of embodiment 39, wherein forming a plurality of channels in the first membrane comprises:

[0452] (a) heating the first film at a predetermined pressure and a predetermined temperature for a predetermined time; and

[0453] (b) molding the plurality of channels using a mold.

[0454] 41. The method of embodiment 40, wherein the fusing of the second film to the first film is performed in the mold.

[0455] 42. The method of embodiment 41, wherein the mold comprises a positive mold.

[0456] 43. The method of embodiment 41, wherein the mold comprises a negative mold.

[0457] 44. The method of embodiment 40, wherein the predetermined temperature is from about 100°C to about 600°C.

[0458] 45. The method of embodiment 40, wherein the predetermined pressure is about 2 pounds per square inch (psi) to about 140 psi.

[0459] 46. ​​The method of embodiment 40, wherein the predetermined time is about 3 minutes to about 30 minutes.

[0460] 47. The method of embodiment 40, wherein the predetermined pressure is about 3.5 psi, and wherein the predetermined temperature is about 370°C.

[0461] 48. The method of embodiment 39, wherein forming a plurality of channels in the first membrane and fusing the second membrane to the first membrane comprises:

[0462] (a) placing the first film and the second film in a frame, wherein the first film and the second film are substantially parallel, substantially aligned, and separated by a gap distance; and

[0463] (b) impacting one or more points on the first film with a fusing tool, wherein the fusing tool is heated to a set fusing temperature, and wherein the fusing tool is in contact with the film for a set fusing time during each impact.

[0464] 49. The method of embodiment 48, wherein striking the first film pierces the first film, the second film, or both, and fuses a portion of the first film to the second film.

[0465] 50. The method of embodiment 48, wherein the frame surrounds at least a portion of the outer edges of the first and second films.

[0466] 51. The method of embodiment 48, wherein the gap distance is about 300 μm to about 1,200 μm.

[0467] 52. The method of embodiment 48, wherein the impact contact area of ​​the fusion tool is at least about 0.07 mm2.

[0468] 53. The method of embodiment 48, wherein striking one or more points on the first film with a fusion tool comprises striking each of the one or more points at most about 16 times.

[0469] 54. A method as described in embodiment 53, wherein the impacting one or more points on the first film with a fusion tool includes impacting each of the one or more points 1 to 6 times.

[0470] 55. The method of embodiment 53, wherein the set fusion temperature is about 250°C to about 1,600°C.

[0471] 56. A method as described in embodiment 53, wherein the set fusion time is less than about 1 second.

[0472] 57. The method of embodiment 39, wherein at least one of the first film and the second film is substantially flat.

[0473] 58. The method of embodiment 39 further comprises embossing the first film before forming the plurality of channels in the first film.

[0474] 59. The method of embodiment 39, further comprising laser ablating a portion of the first film and the second film within the plurality of channels.

[0475] 60. The method of embodiment 59, wherein the laser ablation removes a fused portion of the first film and the second film to form an opening.

[0476] 61. The method of embodiment 60, wherein the concentricity of the opening relative to the channel is at most 25% of the diameter of the channel.

[0477] 62. A method as described in embodiment 39, wherein at least one of the first membrane and the second membrane comprises PVDF, PTFE, ePTFE, PCL, PE / PES, PP, PS, PMMA, PLGA, PLLA or any combination thereof.

[0478] 63. The method of embodiment 39, further comprising coating the device with a hydrophilic polymer.

[0479] 64. The method of embodiment 39, wherein the first film is sintered.

[0480] 65. A method as described in embodiment 39, wherein the second film is not sintered.

[0481] 66. The method of embodiment 39, wherein the second film and the first film are fused using a fusion peel force of at least about 0.2N.

[0482] 67. A method comprising:

[0483] (a) contacting tissue of a diabetic or prediabetic subject with a device comprising a population of insulin-secreting cells, wherein the device comprises:

[0484] (i) a first membrane having a first surface including a plurality of channels and a plurality of second surfaces opposite the first surface; and

[0485] (ii) a second film, the second film being opposite to the first film and attached to the plurality of the second surfaces of the first film;

[0486] wherein the first membrane and the second membrane form a closed compartment having a surface area to volume ratio of at least about 40 cm -1 ,and

[0487] wherein the enclosed compartment provides a volume for containing cells within the device; and

[0488] (b) releasing insulin from the population of insulin-secreting cells in response to elevated blood glucose levels in the diabetic subject, wherein the elevated blood glucose levels are higher than blood glucose levels in non-diabetic subjects.

[0489] 68. A method as described in any of embodiments 67, wherein the insulin-secreting cell population releases an amount of insulin sufficient to reduce blood glucose levels in the diabetic or prediabetic subject.

[0490] 69. The method of embodiment 68, wherein the release of insulin is stopped when the blood glucose level in the diabetic subject decreases to a normal level.

[0491] 70. The method of embodiment 69, wherein the release of insulin is resumed when the population of insulin-secreting cells is re-exposed to elevated blood glucose levels in the diabetic subject.

[0492] 71. A method as described in embodiment 70, wherein the insulin-secreting cell population is a stem cell-derived cell population.

[0493] 72. The method of embodiment 71, wherein the insulin-secreting cell population is capable of glucose-stimulated insulin secretion (GSIS).

[0494] 73. The method of embodiment 68, wherein at least one of the first membrane and the second membrane is semi-permeable.

[0495] 74. The method of embodiment 73, wherein the semi-permeability of the first membrane, the second membrane, or both is configured to protect the cells from immune attack.

[0496] 75. The method of embodiment 74, wherein the semi-permeability of the first membrane, the second membrane, or both is configured to protect the cells from immune attack in the absence of immunosuppressive therapy.

[0497] 76. The method of embodiment 68, wherein at least one of the first membrane and the second membrane is configured to achieve vascularization of the cells within the device.

[0498] 77. The method of embodiment 76, wherein at least one of the first membrane and the second membrane is configured to achieve vascularization of the cells within the device in the absence of immunosuppressive therapy.

Claims

1. A cell containing device, the cell containing device include: (a) a first membrane having a first surface including a plurality of channels and a plurality of second surfaces opposite the first surface; as well as (b) a second film, the second film being opposite to the first film and attached to the plurality of the second surfaces of the first film; wherein the first membrane and the second membrane form a closed compartment having a surface area to volume ratio of at least about 40 cm -1 ,and Wherein the enclosed compartment provides a volume for containing cells within the device.

2. The device of claim 1, wherein the compartment comprises a single continuous open space.

3. The device of claim 1, wherein the volume is about 8 μl to about 1,000 μl.

4. The device of claim 1, wherein at least one of the length and width of the device is about 0.25 cm to about 3 cm.

5. The device of claim 1, having a thickness of at least about 300 μm.

6. The device of claim 1, wherein the plurality of channels are substantially perpendicular to the first membrane.

7. The device of claim 1, wherein the plurality of channels are arranged in a linear array.

8. The device of claim 1, wherein the plurality of channels are arranged in an annular array.

9. The device of claim 1, wherein the plurality of channels have an average diameter of about 400 μm to about 3,000 μm.

10. The device of claim 9, wherein the diameter is measured at the narrowest point in the plurality of channels.

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