Cell containment device

Abstract

To provide a cell housing device.SOLUTION: The present disclosure provides cell housing devices and methods of manufacturing devices having arrays of channels that increase a surface area to volume ratio. In a certain aspect, described herein, a cell housing device, comprises: a first membrane having a first surface comprising a plurality of channels, and a plurality of second surfaces opposing the first surface; and a second membrane opposite and attached to the plurality of the second surfaces of the first membrane, the first membrane and the second membrane forming an enclosed compartment having a surface area to the volume ratio of at least about 40 cm-1, and the enclosed compartment providing a volume for housing a cell within the device.SELECTED DRAWING: None

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the benefit of U.S. Provisional Application No. 62 / 565,962, filed Sep. 29, 2017, and U.S. Provisional Application No. 62 / 671,297, filed May 14, 2018. These documents are hereby incorporated by reference herein.

Background Art

[0002] For treating metabolic disorders such as diabetes, therapeutic devices for delivering biological products can be used. The therapeutic devices may be implantable to provide biological products such as insulin over an extended period of time. These devices may include a cell - containing device and a matrix contained within the cell - containing device. The matrix may contain cells that produce biological products. As the size of the matrix increases, the availability of oxygen and other nutrients may further decrease away from the edge surface of the matrix, and there may be regions within the matrix where the concentration of oxygen and nutrients is low or zero. These regions of low or zero oxygen and nutrient concentration may not be able to support cell viability and biological product synthesis in the matrix. Spatial limitations in transporting oxygen, nutrients, and other agents may limit the size of the device to dimensions where oxygen, nutrients, and other agents can reach the cells. Thus, it can be useful to improve mass transport to the interior regions of such devices and to the interior regions of the matrix contained within the cell - containing device.

Summary of the Invention

Means for Solving the Problems

[0003] The present disclosure generally relates to medical devices and methods. In various aspects, the present disclosure provides medical devices that include a cell - containing device, related devices, and methods of manufacturing and using such devices.

[0004] In certain embodiments, described herein is a cell containment device comprising a first membrane having a first surface that includes a plurality of channels and a plurality of second surfaces that face and are opposite the first surface, and a second membrane that faces and is attached to the plurality of second surfaces of the first membrane, wherein the first membrane and the second membrane form an enclosed compartment having a surface area to volume ratio of at least about 40 cm -1 and the enclosed compartment provides a volume for containing cells within the device.

[0005] In some embodiments, the compartment includes a single continuous open space. In some embodiments, the volume is from about 8 μL to about 1,000 μL. In some embodiments, the device has at least one of a length and a width from about 0.25 cm to about 3 cm. In some embodiments, the device has a thickness of 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 a polar array. In some embodiments, the channels have an average diameter from about 400 μm to about 3,000 μm. In some embodiments, the diameter is measured at the narrowest point within the channel. In some embodiments, the center of each channel is separated from the center of another channel by a distance from about 75 μm to about 500 μm. In some embodiments, the channels have a height to diameter ratio of at least about 0.2. In some embodiments, the device has a channel number per area along a cross-section of about 50 / cm 2Greater than. In some embodiments, at least one of the first membrane and the second membrane includes a plurality of nodes interconnected by a plurality of small fibers. In some embodiments, at least one of the first membrane and the second membrane includes PVDF, PTFE, ePTFE, PCL, PE / PES, PP, PS, PMMA, PLGA, PLLA, or any combination thereof. In some embodiments, the device further includes an opening through the first membrane and the second membrane within the channel. In some embodiments, the opening is concentric with the channel and is at most 25% of the diameter of the channel. In some embodiments, the device further includes a frame configured to receive the device. In some embodiments, the frame is configured to receive a plurality of cell-containing devices. In some embodiments, the frame includes a flexible mechanism configured to prevent buckling of the cell-containing device. In some embodiments, the device further includes a cell population. In some embodiments, the cell population is an insulin-secreting population. In some embodiments, the cell population is stem cell-derived cells capable of glucose-stimulated insulin secretion (GSIS). In some embodiments, the device further includes a coating comprising a hydrophilic polymer. In some embodiments, the device has an insulin diffusion coefficient of about 2x10^-6 cm 2 / s to about 1x10^-5 cm 2 / s. In some embodiments, the device has a maximum insulin diffusion distance of less than about 150 μm. In some embodiments, the first and second membranes are fused with a fusion detachment force of at least about 0.4 N. In some embodiments, at least one of the first and second membranes 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 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. In some embodiments, at least one of the first and second membranes is configured to enable angiogenesis of the cells within the device. In some embodiments, at least one of the first and second membranes is configured to enable angiogenesis of the cells within the device in the absence of immunosuppressive therapy.

[0006] Another aspect provided herein is a cell containment device comprising a first membrane having a first surface including a plurality of channels and a plurality of second surfaces opposite the first surface, and a second membrane facing and attached to the plurality of second surfaces of the first membrane, wherein the first and second membranes form an enclosed compartment, the enclosed compartment providing a volume for containing from 1 million to 1 billion insulin-producing cells within the device, and the membranes are a cell containment device that allows diffusion of insulin from the device while retaining insulin-producing cells within the device.

[0007] Another aspect provided herein is a composition comprising insulin-producing cells and a device containing the insulin-producing cells, wherein the device, when implanted into an individual, releases insulin while retaining the insulin-producing cells therein and facilitates tissue angiogenesis within and around the device. In some embodiments, the individual is not administered an immunosuppressant during implantation of the device or angiogenesis. In some embodiments, the device contains from 1 million to 1 billion insulin-producing cells. In some embodiments, the device is at least about 300 μm thick. In some embodiments, the device comprises a membrane including a plurality of nodes interconnected by a plurality of small fibers.

[0008] In another aspect, described herein is a method of manufacturing a cell-containing device, the method comprising providing a first membrane having a first surface and an opposing second surface, forming a plurality of channels within the first surface of the first membrane, and fusing a second membrane to the second surface of the first membrane to form a compartment for containing cells between the second surface of the first membrane and the second membrane.

[0009] In some embodiments, forming a plurality of channels within the first membrane includes heating the first membrane at a predetermined time, a predetermined pressure, and a predetermined temperature, and forming the plurality of channels using a mold. In some embodiments, fusing the second membrane to the first membrane is performed within the mold. In some embodiments, the mold includes a positive mold. In some embodiments, the mold includes a negative mold. In some embodiments, the predetermined temperature is from about 100 °C to about 600 °C. In some embodiments, the predetermined pressure is from about 2 pounds per square inch (psi) to about 140 psi. In some embodiments, the predetermined time is from about 3 minutes to about 30 minutes. In some embodiments, the predetermined pressure is about 3.5 psi and the predetermined temperature is about 370 °C. In some embodiments, forming a plurality of channels within the first membrane and fusing the second membrane to the first membrane includes placing the first membrane and the second membrane within a frame, wherein the first membrane and the second membrane are substantially parallel, substantially aligned, and separated by a gap distance; placing, and using a fusion tool to dot one or more points on the first membrane, wherein the fusion tool is heated to a set fusion temperature and the fusion tool contacts the membrane for a set fusion time between each dot; and dotting. In some embodiments, dotting the first membrane fuses a portion of the first membrane to the second membrane by penetrating the first membrane, the second membrane, or both. In some embodiments, the frame surrounds at least a portion of the outer edges of the first membrane and the second membrane. In some embodiments, the gap distance is from about 300 μm to about 1,200 μm. In some embodiments, the fusion tool has a dot contact area of at least about 0.07 mm 2It is. In some embodiments, dotting one or more points on the first membrane using a fusion tool includes dotting each of the one or more points up to about 16 times. In some embodiments, dotting one or more points on the first membrane using a fusion tool includes dotting each of the one or more points 1 to 6 times. In some embodiments, the set fusion temperature is from 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 membrane and the second membrane is substantially flat. In some embodiments, the method further includes embossing the first membrane before forming a plurality of channels in the first membrane. In some embodiments, the method further includes laser ablating a portion of the first membrane and the second membrane within the plurality of channels. In some embodiments, the fusion portion between the first membrane and the second membrane is removed by laser ablation to form an opening. In some embodiments, the opening has a concentricity with respect to the channel of up to 25% of the diameter of the channel. In some embodiments, at least one of the first membrane and the second membrane includes PVDF, PTFE, ePTFE, PCL, PE / PES, PP, PS, PMMA, PLGA, PLLA, or any combination thereof. In some embodiments, the method further includes coating the device with a hydrophilic polymer. In some embodiments, the first membrane is sintered. In some embodiments, the second membrane is not sintered. In some embodiments, the second membrane and the first membrane are fused with a fusion peel force of at least about 0.2 N.

[0010] Another aspect provided herein is a method of contacting tissue of a subject with diabetes or prediabetes with an apparatus that includes a population of insulin-secreting cells, the apparatus including a first membrane having a first surface that includes a plurality of channels and a plurality of second surfaces that face and are opposed to the first surface, and a second membrane that faces and is attached to the plurality of second surfaces of the first membrane, wherein the first membrane and the second membrane form an enclosed compartment having a surface area to volume ratio of at least about 40 cm -1 and that provides a volume for containing cells within the apparatus, the contacting, and releasing insulin from the population of insulin-secreting cells in response to an increase in blood glucose level in a subject with diabetes, the increased glucose level being higher than the blood glucose level in a subject without diabetes.

[0011] In some embodiments, the insulin-secreting cell population releases an amount of insulin sufficient to lower blood glucose levels in a subject with diabetes or prediabetes. In some embodiments, releasing insulin ceases when the blood glucose level in a subject with diabetes has dropped to a normal level. In some embodiments, releasing insulin resumes when the insulin-secreting cell population is again exposed to an increase in blood glucose level in a subject with diabetes. In some embodiments, the insulin-secreting cell population is a stem cell-derived cell population. In some embodiments, the insulin-secreting cell population is capable of glucose-stimulated insulin secretion (GSIS). In some embodiments, at least one of the first membrane and the second membrane 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 membrane and the second membrane is configured to permit angiogenesis of the cells within the device. In some embodiments, at least one of the first membrane and the second membrane is configured to permit angiogenesis of the cells within the device in the absence of immunosuppressive therapy.

[0012] In certain aspects, described herein is a cell containment device comprising a first surface defining an outer surface of the device and having a surface area, and a second surface opposing the first surface and defining an inner surface of the device, and a compartment enclosed within the second surface and providing a volume for containing cells within the device, wherein the surface area to volume ratio is 50 cm -1The above is the cell-containing device. In some embodiments, the device includes a plurality of channels that extend through the cross-section of the device. In some embodiments, each channel of the plurality of channels has a diameter of 400 μm or more. In some embodiments, the diameter is measured at the narrowest point within the channel. In some embodiments, each channel of the plurality of channels is separated from each other by a distance not exceeding 450 μm. In some embodiments, each channel of the plurality of channels has a height-to-diameter ratio of 0.2 or more. In some embodiments, the number of channels per unit area measured along the cross-section of the device is greater than 50 / cm 2 . In some embodiments, the number of channels per unit area measured along the cross-section of the device is greater than 100 / cm 2 . In some embodiments, the surface area-to-volume ratio is greater than 80 cm -1 . In some embodiments, the surface area-to-volume ratio is greater than 100 cm -1 . In some embodiments, the surface area-to-volume ratio is greater than 120 cm -1 . In some embodiments, the device includes a single continuous open space having a volume. In some embodiments, the first surface or the second surface includes a plurality of nodes interconnected by a plurality of small fibers. In some embodiments, the device has a thickness measured along the cross-section of the device greater than 300 μm. In some embodiments, the first surface or the second surface includes PVDF, PTFE, ePTFE, PCL, PE / PES, PP, PS, PMMA, PLGA, or PLLA. In some embodiments, the device further includes a frame configured to receive the device. In some embodiments, the frame is configured to receive a plurality of cell-containing devices. In some embodiments, the frame includes a flexible mechanism to prevent buckling of the cell-containing device. In some embodiments, the device further includes a cell population. In some embodiments, the cell population is an insulin-secreting population. In some embodiments, the cell population is stem cell-derived cells capable of glucose-stimulated insulin secretion (GSIS).

[0013] In some embodiments, the device further includes a coating with a hydrophilic polymer. In some embodiments, the volume for containing cells is inversely proportional to at least one of the diameters of the plurality of channels and the number of channels per unit area of the device. In some embodiments, the surface area to volume ratio of the device is directly proportional to at least one of the diameters of the plurality of channels and the number of channels per unit area of the device. In some embodiments, the surface area to volume ratio of the device can increase mass transport into and / or out of the device. In other embodiments, described herein is a cell containment device comprising a base, an upper surface opposite the base, a height extending from the base to the upper surface along a cross-section of the device, the height being longer than 300 μm, a compartment for containing cells, the compartment being enclosed between the base and the upper surface, and a plurality of channels extending along the cross-section of the device, wherein the maximum oxygen diffusion distance of the device is less than 150 μm. In some embodiments, the device has a height greater than 600 μm. In some embodiments, the base is substantially flat. In some embodiments, each channel of the plurality of channels has a diameter of 400 μm or more. In some embodiments, each channel of the plurality of channels is separated from each other by a distance not exceeding 450 μm. In some embodiments, each channel of the plurality of channels has a diameter of 400 μm or more. In some embodiments, the diameter is measured at the narrowest point within the channel. In some embodiments, each of the plurality of channels is separated from each other by a distance not exceeding 450 μm. In some embodiments, each of the plurality of channels has a height to diameter ratio of 0.2 or more. In some embodiments, the number of channels per unit area measured along the cross-section of the device is greater than 50 / cm 2 or more. In some embodiments, the number of channels per unit area measured along the cross-section of the device is 100 / cm 2Larger. In some embodiments, the device includes a single continuous compartment for containing cells. In some embodiments, the first surface or the second surface includes a plurality of nodes interconnected by a plurality of small fibers. In some embodiments, the first surface or the second surface includes PVDF, PTFE, ePTFE, PCL, PE / PES, PP, PS, PMMA, PLGA, or PLLA. In some embodiments, the device further includes a frame configured to receive the device. In some embodiments, the frame is configured to receive a plurality of cell-containing devices. In some embodiments, the frame includes a flexible mechanism to prevent buckling of the cell-containing device. In some embodiments, the device further includes a cell population. In some embodiments, the cell population is an insulin-secreting population. In some embodiments, the cell population is stem cell-derived cells capable of glucose-stimulated insulin secretion (GSIS). In some embodiments, the device further includes a coating with a hydrophilic polymer. In some embodiments, the volume for containing cells is inversely proportional to at least one of the diameter of the plurality of channels and the number of channels per area of the device. In some embodiments, the surface area-to-volume ratio of the device is directly proportional to at least one of the diameter of the plurality of channels and the number of channels per area of the device. In some embodiments, the surface area-to-volume ratio of the device is directly proportional to at least one of the diameter of the plurality of channels and the number of channels per area of the device. In some embodiments, by increasing the surface area-to-volume ratio of the device, mass transport into and / or out of the device can be increased.

[0014] In another aspect, described herein is a method of manufacturing a cell-containing device, the method comprising: providing a first membrane; adjusting the temperature and / or pressure surrounding the first membrane to a predetermined value; deforming the first membrane; and fusing a second membrane to the first membrane, wherein a compartment for containing cells is defined by a compartment between the first membrane and the second membrane. In some aspects, the predetermined value is less than 170 degrees Celsius (°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 5 psi. In some aspects, deforming the first membrane includes pushing down a portion of the first membrane with a tool. In some aspects, the tool includes a substantially flat surface configured to be parallel to the first membrane and a plurality of protrusions on the surface configured to push down a portion of the first membrane. In some aspects, each of the plurality of protrusions includes a cylinder. In some aspects, the tool includes a tip portion having a contact area at the free end. In some aspects, the contact area is 0.07 mm 2The above is the case. In some embodiments, the fusion is performed using the tip, and the tip presses the first and second membranes and brings them into contact with each other for a predetermined time. In some embodiments, the deformation and fusion are performed in one step using the tip, the tip contacts the first membrane, deviates from the first membrane, moves in a vertical direction toward the second membrane, presses the first and second membranes, and brings them into contact with each other for a predetermined time. In some embodiments, the tip is adjusted to a temperature of a predetermined value. In some embodiments, the tip presses with a pressure of a predetermined value. In some embodiments, the predetermined time is 1 second or more. In some embodiments, the cylinder has a diameter of 300 μm or more. In some embodiments, the cylinder has a height of 300 μm or more. In some embodiments, deforming the first membrane is performed without causing cracks in the membrane. In some embodiments, deforming the first membrane includes forming a plurality of features on the membrane. In some embodiments, each of the plurality of features has a diameter of 300 μm or more. In some embodiments, each of the plurality of features has a depth of 300 μm or more. In some embodiments, the method further includes adjusting the temperature and / or pressure surrounding the first membrane to control the characteristics of the features. In some embodiments, increasing the temperature and / or pressure surrounding the first membrane increases the depth of the features. In some embodiments, fusing the second membrane to the first membrane includes fusing the second membrane and the first membrane into one continuous layer. In some embodiments, fusing the second membrane to the first membrane begins at a temperature and / or pressure having a second predetermined value. In some embodiments, the second predetermined value is less than 230 degrees Celsius (°C). In some embodiments, the second membrane is substantially flat. In some embodiments, after deformation, the first membrane is embossed. In some embodiments, after fusion, the device has a substantially flat surface and an embossed surface facing the substantially flat surface.In some embodiments, the method further includes forming a channel that traverses through the device by removing the fused portion of the first membrane and the second membrane by laser ablation. In some embodiments, the method further includes attaching the device to a frame. In some embodiments, the method further includes implanting the device on the frame into a subject. In some embodiments, the method further includes encapsulating cells within a compartment. In some embodiments, the method further includes implanting the device into a subject. In some embodiments, the first membrane or the second membrane includes PVDF, PTFE, ePTFE, PCL, PE / PES, PP, PS, PMMA, PLGA, or PLLA. In some embodiments, the method further includes coating the device with a hydrophilic polymer. In some embodiments, the first membrane is sintered. In some embodiments, the second membrane is not sintered.

[0015] In another aspect, described herein is a cell containment device comprising a base, an upper surface opposite the base, a height reaching from the base to the upper surface along a cross-section of the device and being 300 μm or less, and a compartment for containing cells, the compartment being enclosed between the base and the upper surface. In some aspects, the height is less than 250 μm. In some aspects, the base includes a sintered membrane. In some aspects, the upper surface includes a sintered membrane. In some aspects, the base includes a coated membrane, and the coating increases the hydrophilicity of the membrane. In some aspects, the device includes at least one fused dot, the dot including fusing a part of the base and a part of the upper surface corresponding to the part of the base, and the dot being configured to limit a change with respect to the height. In some aspects, the dot has a diameter of about 0.5 mm to about 3 mm. In some aspects, the dot has a diameter of at least about 0.5 mm. In some aspects, the dot has a maximum diameter of about 3 mm.In some embodiments, the dots have a diameter of from about 0.5 mm to about 0.75 mm, from about 0.5 mm to about 1 mm, from about 0.5 mm to about 1.25 mm, from about 0.5 mm to about 1.5 mm, from about 0.5 mm to about 1.75 mm, from about 0.5 mm to about 2 mm, from about 0.5 mm to about 2.25 mm, from about 0.5 mm to about 2.5 mm, from about 0.5 mm to about 2.75 mm, from about 0.5 mm to about 3 mm, from about 0.75 mm to about 1 mm, from about 0.75 mm to about 1.25 mm, from about 0.75 mm to about 1.5 mm, from about 0.75 mm to about 1.75 mm, from about 0.75 mm to about 2 mm, from about 0.75 mm to about 2.25 mm, from about 0.75 mm to about 2.5 mm, from about 0.75 mm to about 2.75 mm, from about 0.75 mm to about 3 mm, from about 1 mm to about 1.25 mm, from about 1 mm to about 1.5 mm, from about 1 mm to about 1.75 mm, from about 1 mm to about 2 mm, from about 1 mm to about 2.25 mm, from about 1 mm to about 2.5 mm, from about 1 mm to about 2.75 mm, from about 1 mm to about 3 mm, from about 1.25 mm to about 1.5 mm, from about 1.25 mm to about 1.75 mm, from about 1.25 mm to about 2 mm, from about 1.25 mm to about 2.25 mm, from about 1.25 mm to about 2.5 mm, from about 1.25 mm to about 2.75 mm, from about 1.25 mm to about 3 mm, from about 1.5 mm to about 1.75 mm, from about 1.5 mm to about 2 mm, from about 1.5 mm to about 2.25 mm, from about 1.5 mm to about 2.5 mm, from about 1.5 mm to about 2.75 mm, from about 1.5 mm to about 3 mm, from about 1.75 mm to about 2 mm, from about 1.75 mm to about 2.25 mm, from about 1.75 mm to about 2.5 mm, from about 1.75 mm to about 2.75 mm, from about 1.75 mm to about 3 mm, from about 2 mm to about 2.25 mm, from about 2 mm to about 2.5 mm, from about 2 mm to about 2.75 mm, from about 2 mm to about 3 mm, from about 2.25 mm to about 2.5 mm, from about 2.25 mm to about 2.75 mm, from about 2.25 mm to about 3 mm, from about 2.5 mm to about 2.75 mm, from about 2.5 mm to about 3 mm, or from about 2.75 mm to about 3 mm. In some embodiments, the dots have a diameter of 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 embodiments, the dots are spaced apart from any other fused dots by at least 3 mm. In some embodiments, the dots are formed using an adhesive disposed between a portion of the base and a portion of the top surface.In some embodiments, the volume of the compartment is inversely proportional to at least one of the diameter of the dots and the number of dots per unit area of the device. In some embodiments, the ratio of the surface area of the device to the volume of the device is directly proportional to at least one of the diameter of the dots and the number of dots per unit area of the device. In some embodiments, by increasing the surface area-to-volume ratio of the device, mass transport into and / or out of the device can be increased.

[0016] In other embodiments, described herein is a method comprising: a) contacting tissue of a subject with diabetes with a device comprising a population of insulin-secreting cells, the device comprising a first surface defining an outer surface of the device and having a surface area, a second surface opposite the first surface and defining an inner surface of the device, and a compartment enclosed within the second surface and providing a volume for containing cells within the device, the surface area-to-volume ratio being 50 cm -1 or greater; and b) releasing insulin from the population of insulin-secreting cells in response to an increase in blood glucose level in the subject with diabetes, the elevated glucose level being higher than the blood glucose level in a non-diabetic subject. In some embodiments, the population of insulin-secreting cells releases an amount of insulin sufficient to lower the blood glucose level in the subject with diabetes. In some embodiments, releasing insulin ceases when the blood glucose level in the subject with diabetes has dropped to a normal level. In some embodiments, releasing insulin resumes when the population of insulin-secreting cells is again exposed to an increase in blood glucose level in the subject with diabetes. In some embodiments, the population of insulin-secreting cells is a stem cell-derived cell population. In some embodiments, the population of insulin-secreting cells is capable of glucose-stimulated insulin secretion (GSIS).

[0017] In another aspect, described herein is a method comprising: a) contacting tissue of a subject with diabetes with an apparatus comprising an insulin-secreting cell population, the apparatus comprising a base, an upper surface facing the base, a height reaching from the base to the upper surface along a cross-section of the apparatus and being longer than 300 μm, a compartment for containing cells, the compartment being enclosed between the base and the upper surface, and a plurality of channels extending along the cross-section of the apparatus, wherein a maximum oxygen diffusion distance of the apparatus is less than 150 μm; and b) releasing insulin from the insulin-secreting cell population in response to an increase in blood glucose level in the subject with diabetes, the increased glucose level being higher than a blood glucose level in a non-diabetic subject. In some aspects, the insulin-secreting cell population releases an amount of insulin sufficient to lower the blood glucose level in the subject with diabetes. In some aspects, releasing insulin ceases when the blood glucose level in the subject with diabetes has dropped to a normal level. In some aspects, releasing insulin resumes when the insulin-secreting cell population is re-exposed to an increase in blood glucose level in the subject with diabetes. In some aspects, the insulin-secreting cell population is a stem cell-derived cell population. In some aspects, the insulin-secreting cell population is capable of glucose-stimulated insulin secretion (GSIS).

[0018] In another aspect, described herein is a method comprising: a) contacting tissue of a subject with diabetes with an apparatus comprising an insulin-secreting cell population, the apparatus comprising a base, an upper surface opposite the base, a height reaching from the base to the upper surface along a cross-section of the apparatus and being 300 μm or less, and a compartment for containing cells, the compartment being enclosed between the base and the upper surface; and b) releasing insulin from the insulin-secreting cell population in response to an increase in blood glucose level in the subject with diabetes, the increased glucose level being higher than the blood glucose level in a non-diabetic subject. In some aspects, the insulin-secreting cell population releases an amount of insulin sufficient to lower the blood glucose level in the subject with diabetes. In some aspects, releasing insulin ceases when the blood glucose level in the subject with diabetes has dropped to a normal level. In some aspects, releasing insulin resumes when the insulin-secreting cell population is re-exposed to an increase in blood glucose level in the subject with diabetes. In some aspects, the insulin-secreting cell population is a stem cell-derived cell population. In some aspects, the insulin-secreting cell population is capable of glucose-stimulated insulin secretion (GSIS).

[0019] Incorporation by reference All publications, patents, and patent applications mentioned herein are hereby 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.

[0020] The novel features of the disclosure are set forth in the appended claims. The features and advantages of the disclosure will be better understood by reference to the following detailed description of illustrative embodiments using the principles of the disclosure and the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0021]

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DETAILED DESCRIPTION OF THE INVENTION

[0022] The present disclosure generally relates to medical devices and methods. The medical devices may include cell-containing devices, devices related thereto, and methods of manufacturing and using such devices. The devices may serve to improve mass transport between environments that are external and internal to the device.

[0023] In some cases, the cell containment device may include a high surface area to volume ratio. The high surface area to volume ratio may enable the device to achieve improved mass transport into and / or out of the device, which may enhance the effectiveness of delivering nutrients to the cells within the device. In some cases, the cell containment device may include a first surface. The first surface may define the outer surface of the device and may have a surface area. The cell containment device may also include a second surface opposite the first surface, which defines the inner surface of the device. The cell containment device may also include a compartment enclosed within the second surface, which provides a volume for containing cells within the device. The device may include a single continuous open space having a volume. The first surface or the second surface of the device may include a plurality of nodes interconnected by a plurality of small fibers. The device may also include a plurality of channels that extend through the cross-section of the device. The channels may provide a high surface area to volume ratio for the cell containment device. Each channel may have a diameter of 400 μm or more, and the diameter may be measured at the narrowest point within the channel. The plurality of channels may be separated from each other by a distance not exceeding 450 μm. The device may have a thickness greater than 250 μm when measured along the cross-section of the device. In some cases, the channels may be configured such that the thickness of the cell containment device does not pose a problem for mass transport into and / or out of the device (e.g., for nutrients). 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 further include a frame, which is configured to receive one or more cell containment devices. The frame may include a flexible mechanism to prevent buckling of the cell containment device. The device may include a coating with a hydrophilic polymer. The volume for containing cells may be 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. The surface area to volume ratio of the device may be directly proportional to at least one of the diameter of the plurality of channels and the number of channels per unit area of the device.The surface area to volume ratio of the device may allow for greater mass transport into and / or out of the device.

[0024] In some cases, the cell containment device may have a short oxygen diffusion distance. This short oxygen diffusion distance may be independent of the dimensions of the cell containment device (e.g., its thickness). For example, the cell containment device may include a base and an upper surface opposite the base. A compartment for containing cells may be enclosed between the base and the upper surface, and in some cases, may rely on mass transport from outside the cells to access nutrients for life support. The height reaching from the base to the upper surface along a cross-section of the device may be a high value (e.g., greater than 300 μm). Nevertheless, the oxygen diffusion distance to the device may be less than 150 μm. In some cases, this may be enabled by various channels or lumens of the device. The channels may, in some cases, extend across the cross-section of the device and may allow for a short oxygen diffusion distance in the device independent of its thickness. The base may be substantially flat.

[0025] The cell-containing device described in this specification may be manufactured using two membranes. In one step, the first membrane may be prepared using various materials described in this specification. In some cases, the first membrane may be provided within a chamber in which the temperature and / or pressure may be set and / or adjusted to a predetermined value. The first membrane may be deformed (e.g., to form a compartment or volume for containing cells). Thereafter, the second membrane may be fused to the first membrane to form a compartment. Optionally, an aperture may be formed within a channel of the device to enable a continuous passage through the cell-containing device using various means such as a laser. In some cases, the temperature and / or pressure may be adjusted to deform the first membrane or to fuse the first membrane to the second membrane. The temperature and / or pressure used in each of these processes may be an essential part of manufacturing the cell-containing device. By adjusting the temperature and / or pressure in the deformation or fusion region, the characteristics of the feature may be controlled. In some cases, the depth of the feature may be increased by increasing the temperature and / or pressure in the deformation or fusion region. In some cases, after deformation, the first membrane may be embossed. In some cases, after fusion, the device may have a substantially flat surface and an embossed surface facing the substantially flat surface. Shown in FIG. 1 is a high-magnification image of a cell-containing device formed using a PVDF membrane and having a regularly spaced array of apertures within a channel of the device. Deforming the membrane may further (or alternatively) include fibrillation. Nodes from one membrane unwind and interact with fibrils from another membrane to form entanglements and seals.

[0026] As described above, when cells that produce various biological products are encapsulated in a device, it may be very useful, for example, for delivering treatment. In this specification, the device may be referred to as a cell-containing device, and a matrix may be contained inside the device. The matrix can be a biological material in the internal space of the cell-containing device. The matrix may include a hydrogel, a porous sponge, an electrospun fiber, a polymer material, or other porous biocompatible materials. The matrix may further include growth factors, nutrients, or other agents that enhance cell activity and the synthesis of biological products. The matrix may include cells or other protein expression systems (e.g., cell-free expression systems) that enable the production of biological products. As the thickness of the matrix increases, the availability of oxygen and other nutrients may further decrease at locations far from the edge surface of the matrix. For example, this may occur when the transport of oxygen or nutrients mainly relies on passive transport or diffusion. As a result, very little or no nutrients may be delivered to regions within the matrix far from the surface. The availability of nutrients may further sharply decrease due to consumption by cells as the nutrients pass through the matrix. The decrease in nutrients within the matrix can be a problem for cells or systems with high demand for oxygen or other nutrients. Since cells become very active after stimulation, the oxygen consumption by cells may increase from the basal state when there is stimulation. The decrease in the availability of nutrients important for cell viability and activity may limit the increase in the size of the matrix and the cell-containing device into which the matrix is placed. The limitation of mass transport within the matrix may limit the expansion of the device size to increase the output of biological products. Generally, reducing the cell density within the matrix to reduce consumption may not be desirable because it is unrealistic to increase the matrix dimensions to offset the decrease in cell density.

[0027] The design of the cell containment device may be adjusted to control the surface area to volume (SA:V) ratio of the device. In some cases, the design may be provided to increase the SA:V ratio of the current device. Increasing the SA:V ratio may help improve the transport (e.g., of nutrients) into the interior region of the device. One such design for increasing the SA:V ratio may be to incorporate channels and / or other geometric shapes through the matrix and the cell containment device. In some cases, the channels may help increase the SA:V ratio of the device and the matrix contained within the device. The channels may include through-holes or lumens that go completely from one side of the cell containment device to the opposite side. The channels may be arranged in a pattern or provided without a distinct pattern. As an example, the channels may be provided in an array pattern with a predetermined spacing between the channels. Alternatively, the channels may be provided in a random pattern.

[0028] Including channels in the cell containment device may make it easier to scale up the size of the device compared to a device without channels. Illustrated in Figure 3 are zones of various oxygen pressures within a two-layer flat sheet and a channel array device. As shown in Figure 3, a device 300 (also referred to herein as a channel array device) with a channel array 301 can have channels that go through the matrix and the cell containment device. The cell containment device with channels 300 may be of any thickness, but may also avoid having zones with low oxygen or nutrients. A conventional two-layer flat sheet may be limited to a thickness of 300 μm or less to avoid zones with low oxygen or nutrients. In the case of a cell containment device with channels, mass transport may be improved and the potential for angiogenesis may be increased compared to a two-layer flat sheet design or other channel-less designs. As a result of these characteristics, it may be easier to scale up the size of the channel array device compared to a device without channels.

[0029] A cell-containing device with channels (also referred to as a channel array device) can increase the surface area to enable mass transport. The increased mass transport can increase the overall diffusion flux across the matrix and the device. This increase can enlarge the dimensions of the matrix and the device and reduce areas with very low or no nutrients. As a result, the maximum oxygen diffusion distance of the device can be less than 150 μm. Optionally, the cell-containing device with channels may enable an increase in angiogenesis potential. Illustrated in FIG. 4 are the surface areas and angiogenesis potential of a device with a flat configuration and a device with channels. FIG. 5 exemplifies an increase in angiogenesis with a device with a flat configuration or a device with channels. Shown in FIG. 6 is a computer-aided design (CAD) rendering of a hexagonal device with increasing channel size but a similar SA:V ratio. As shown in FIG. 4, a device with channels can have a greater number of cells per unit area compared to a device with a flat configuration. This is because sufficient nutrients can be provided to the cells throughout the matrix to support their viability and activity. When implanted in vivo, compared to a device with a flat configuration, the channels allow vascular structures to grow around and through the channels. In a device with a flat configuration, as shown in FIGS. 4 and 5, angiogenesis is limited to the upper and lower surfaces of the device. In some cases, angiogenesis can be increased by the characteristics of the channels and / or the number or density of the channels. For example, the diameter of the channel may be the cross-sectional distance of the channel. The diameter of the channel may be measured at its narrowest point within a single cross-section 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. The channels may be arranged such that the number of channels per unit area is present along the cross-section of the device. Depending on the size, number, and / or density of the channels as described herein, angiogenesis may increase and mass transport of nutrients to the cells may be enhanced. The cell-containing device can protect the matrix and the cells or other contents of the matrix from direct contact with the vascular structures within the channels.The design of the array (e.g., its channel size and spacing) can be altered to change the surface area as well as the vasculature growth pattern and potential.

[0030] The dimensions of the channels can be adjusted to control the volume and SA:V ratio within the cell containment device. In some embodiments, the diameter of the channels can be increased to reduce the volume within the cell containment device. As a result, increasing the diameter of the channels can increase the SA:V ratio with respect to the cell containment device. In some embodiments, the diameter of the channels can be decreased to increase the volume within the cell containment device. Decreasing the diameter of the channels can decrease the SA:V ratio with respect to the cell containment device.

[0031] The matrix can be connected as one or more pieces contained within the cell containment device. The matrix may include a single continuous piece within the cell containment device. Also, the matrix may have channels that pass through its thickness where the channels pass through the cell containment device. The matrix can be a biomaterial within the internal space of the cell containment device. The matrix may include a hydrogel, a porous sponge, electrospun fibers, a polymeric material, or other porous biocompatible materials. The matrix may further include growth factors, nutrients, or other agents that enhance cell activity and the synthesis of biological products. The matrix may include cells or other protein expression systems that enable the production of biological products. Shown in FIG. 7 is a scanning electron micrograph of a cross-section of a cell containment device with a rendering of the cell containment device and an associated interior that may contain a matrix.

[0032] The cell containment device may have various lengths, widths, and heights suitable for its application examples. The length may be the longest dimension on the upper surface of the device. The width may be the dimension perpendicular to the length on the upper surface. The height of the device may also be referred to as the thickness of the device and may reach from the base to the upper surface along the cross-section of the device. In some cases, the length of the cell containment device may be about 0.2 cm or more, 0.5 cm or more, 1.0 cm or more, 1.5 cm or more, 2.0 cm or more, 2.5 cm or more, 3.0 cm or more, 4.0 cm or more, 5.0 cm or more, 6.0 cm or more, 7.0 cm or more, 8.0 cm or more, 9.0 cm or more, 10 cm or more, 20 cm or more, 30 cm or more, 40 cm or more, 60 cm or more, 100 cm or more, 120 cm or more, 150 cm or more, 180 cm or more, or 200 cm or more. In some cases, the width of the cell containment device may be about 0.2 cm or more, 0.5 cm or more, 1.0 cm or more, 1.5 cm or more, 2.0 cm or more, 3.0 cm or more, 4.0 cm or more, or 5.0 cm or more, 6.0 cm or more, 7.0 cm or more, 8.0 cm or more, 9.0 cm or more, or 10 cm or more. In some cases, the height of the cell containment device may be about 100 μm or more, 200 μm or more, 300 μm or more, 400 μm or more, 500 μm or more, 600 μm or more, 700 μm or more, 800 μm or more, 900 μm or more, 1000 μm or more, 0.2 cm or more, 0.3 cm or more, 0.4 cm or more, 0.5 cm or more, 0.6 cm or more, 0.7 cm or more, 0.8 cm or more, 0.9 cm or more, 1.0 cm or more, 2 cm or more, 3 cm or more, 4 cm or more, or 5 cm or more (measured along the cross-section of the device).

[0033] The cell containment device may be designed such that the SA:V ratio is a value appropriate for transporting nutrients and the desired product through the device. In some cases, the SA:V ratio may be 50 cm -1 or more. In other cases, the SA:V ratio may be about 20 cm -1 or more, 40 cm -1 or more, 60 cm -1 or more, 80 cm -1 or more, 100 cm -1 or more, 120 cm -1 or more, 150 cm -1 or more, 200 cm-1 Above, 250 cm -1 Above, 300 cm -1 Above, or any value therebetween may be used. The maximum oxygen diffusion distance of the device may be less than 50 μm, less than 100 μm, less than 150 μm, less than 200 μm, less than 250 μm, less than 300 μm, less than 350 μm, less than 400 μm, less than 450 μm, or less than 500 μm.

[0034] In some cases, the channel 751 of the cell containment device 750 may be substantially cylindrical in shape. Illustrated in FIGS. 3, 6, and 7 are cylindrical channels within the cell containment device. For the purpose of explanation in this specification, mainly cylindrical channels will be described. However, it is understood that the channel may be of any shape. For example, the walls of the channel may be substantially straight, curved, barrel-shaped, or other shapes. In some cases, the cross-sectional area of the channel may vary from the upper surface of the first membrane 711 of the cell containment device to the base of the second membrane 720. In some cases, the device may include a fusion portion 754. For example, the device may have the first membrane 711 fused to the second membrane 720 to provide a compartment 753 within the cell containment device. The compartment 753 may be filled with cells. In some cases, the fusion portion of the channel (e.g., the portion where the first membrane meets the second membrane) may be substantially circular or other shapes. As described elsewhere in this specification, in some cases, an opening 752 may be formed in the fusion portion of the membrane to provide a channel that travels through the cell containment device. The channel may include a cross-sectional distance, or diameter, as described in this specification. Alternatively, the cross-sectional distance of the channel may refer to the diameter when the cross-section of the channel is substantially circular. The diameter of the channel may be measured at its narrowest point within a single cross-section parallel to the plane of the second membrane. Alternatively, the diameter may be measured as the average value of the channel width along the height of the channel or device. Alternatively, the diameter may be measured at its widest point within a single cross-section parallel to the plane of the second membrane. In some cases, the diameter of the channel can be 100 μm or more, 200 μm or more, 300 μm or more, 400 μm or more, 500 μm or more, 600 μm or more, 700 μm or more, 800 μm or more, 900 μm or more, or 1000 μm or more. Optionally, the height of the channel may be proportional to the diameter.In some cases, the height-to-diameter ratio of the channel may be about 0.1 or more, 0.2 or more, 0.3 or more, 0.4 or more, 0.5 or more, 0.6 or more, 0.7 or more, 0.8 or more, 0.9 or more, 1.0 or more, 1.1 or more, 1.2 or more, 1.3 or more, 1.4 or more, 1.5 or more, 1.6 or more, 1.7 or more, 1.8 or more, 1.9 or more, 2.0 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, or 10 or more.

[0035] Multiple channels may proceed through the cross-section of the device. The channels may be arranged such that there is a number of channels per area along the cross-section. In this specification, the number of channels per area may be referred to as the channel density of the device. In some cases, the number of channels per area along the cross-section is about 10 channels / cm 2 or more, 15 channels / cm 2 or more, 20 channels / cm 2 or more, 25 channels / cm 2 or more, 30 channels / cm 2 or more, 35 channels / cm 2 or more, 40 channels / cm 2 or more, 45 channels / cm 2 or more, 50 channels / cm 2 or more, 60 channels / cm 2 or more, 70 channels / cm 2 or more, 80 channels / cm 2 or more, 90 channels / cm 2 or more, 100 channels / cm 2 or more, 110 channels / cm 2 or more, 120 channels / cm 2 or more, 130 channels / cm 2 or more, 140 channels / cm 2 or more, 150 channels / cm 2 or more, 175 channels / cm 2 or more, or 200 channels / cm 2 or more.

[0036] The channels can be spaced apart such that there are no regions receiving low or zero amounts of oxygen or other nutrients important for cell viability and activity. In some cases, the channels can be spaced apart or separated from each other at a distance of about 100 μm or less, 200 μm or less, 300 μm or less, 400 μm or less, 500 μm or less, 600 μm or less, 700 μm or less, 800 μm or less, 900 μm or less, or 1000 μm or less. Optionally, the distance may be measured from the center of one channel to the center of an adjacent channel. In some cases, the cell containment device can have a channel spacing of one distance throughout the device. Alternatively, the cell containment device can have channel spacings of multiple different distances throughout the cell containment device. In some cases, the channels can be arranged in a regular array with a regular channel spacing distance across the device. For example, as illustrated in FIG. 6, the channels may be arranged in a hexagonal array. Alternatively, other arrangements of the channels (e.g., circular, square, etc.) may be provided.

[0037] In some cases, the area of the lumen of the channel can be proportional to the cross-sectional area of the channel. The lumen may be cut from a portion of the fusion region of the device. In some cases, the area of the lumen is about 0% or more, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 99% or more of the cross-sectional area of the channel.

[0038] Multiple lumens may extend through the cross-section of the device. The lumens may be arranged such that there is a number of lumens per area along the cross-section. In this specification, the number of lumens per area may be referred to as the lumen density of the device. In some cases, the number of lumens per area along the cross-section is about 10 lumens / cm 2 or more, 15 lumens / cm 2 or more, 20 lumens / cm 2 or more, 25 lumens / cm 2 or more, 30 lumens / cm 2 or more, 35 lumens / cm 2 or more, 40 lumens / cm 2 or more, 45 lumens / cm2 50 lumens / cm or more 2 60 lumens / cm or more 2 70 lumens / cm or more 2 80 lumens / cm or more 2 90 lumens / cm or more 2 100 lumens / cm or more 2 110 lumens / cm or more 2 120 lumens / cm or more 2 130 lumens / cm or more 2 140 lumens / cm or more 2 150 lumens / cm or more 2 175 lumens / cm or more 2 or 200 lumens / cm or more 2 may be above.

[0039] The channel array device can be applied to various in vivo and in vitro applications. In one example, the device can accommodate cells or expression systems with functions such as pancreatic islet cells within its matrix. The matrix may contain isolated pancreatic islet cells, cells isolated from the pancreas, cells isolated from tissues, stem cells, stem cell-derived cells, induced pluripotent cells, differentiated cells, transformed cells, or an expression system (capable of synthesizing one or more biological products). Optionally, the matrix may contain a second type of cell that supports a first type of cell that synthesizes one or more biological products. The cells can be encapsulated before being placed within the matrix. The cells may be encapsulated within microcapsules or conformally coated. This device can be used to supplement pancreatic islet cell function.

[0040] The design of the channel array within the device can affect the angiogenesis potential or the amount of vasculature that can grow through or around the device. The channels within the device can be designed to increase the angiogenesis potential. Such a device may have improved nutrient transport and a reduced risk of hypoxia to the cells contained therein. Such a device can be made larger in size and can accommodate a larger matrix with cells or other expression systems. With such a device having improved transport, cell viability can be extended from initial generation for over one year. In some cases, cell viability can exceed one month, two months, four months, six months, eight months, ten months, twelve months, fourteen months, sixteen months, eighteen months, twenty months, twenty-two months, twenty-four months, thirty-six months, forty-eight months, or more.

[0041] Changes in the surface area of the channel array device can potentially affect the kinetics of biological products. The biological products may include human cells, animal cells, or genetically modified cells. Due to the increased surface area, the channel array device can increase the production and release of biological products. Illustrated in FIG. 32 is the effect of the device design on insulin response and surface area to volume ratio. As shown in FIG. 32, the channel array device can increase the production and release of biological products compared to a similar-sized device without a channel array. The channel array device may have a larger SA:V ratio than a similar-sized device without a channel array. An increase in the SA:V ratio in the channel array device can potentially increase the flux of biological products compared to a similar-sized device without a channel array. In the case of a matrix with cells having functions such as pancreatic islet cells, an increase in the SA:V ratio can potentially increase the islet equivalent (IEQ) per area. In some cases, as a result, insulin production can be increased and insulin flux can be increased. Illustrated in FIG. 6 is a rendering of a hexagonal channel array device with a similar SA:V ratio and an increasing size. Illustrated in FIG. 18 is a rendering of a cell containment device with similar overall dimensions and varying channel dimensions to achieve different SA:V ratios.

[0042] In some cases, the devices described herein can be assembled and / or mounted on a frame as will be further described later. The frame may be configured to receive one cell-containing device or a plurality of cell-containing devices. Optionally, the device can be mounted on a sub-frame in addition to the frame. The frame can provide flexible support for the devices of the present disclosure (e.g., a channel array device). The frame can prevent unwanted bending of the device. The frame can have one or more flexible mechanisms to prevent buckling of the cell-containing device. The flexible mechanism can prevent buckling of sensitive device regions within the device. The flexible mechanism can have a break that allows the frame to bend. The flexible mechanism can allow the assembly to curve at its implantation site along the tissue. Alternatively, the devices described herein can be used without a frame. For example, the device can be implanted into an individual alone, without using any structural support or frame. Shown in FIG. 50 is a prototype of three cell-containing devices assembled on a frame for human use. The device and the frame holding the device can be formed using materials with a low induced foreign body reaction. The materials for the device and the frame can be selected to reduce inflammation or fibrosis. The device and the frame can be used together with anti-inflammatory or anti-macrophage therapy to further reduce the foreign body reaction.

[0043] The cell-containing device can be manufactured using a simple process. In some cases, the cell-containing device can be manufactured by deforming a first membrane into the shape of a channel array 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, the tip of the channel shape can be used to deform the membrane at the location where the tip contacts the membrane.

[0044] The membrane may include a biocompatible porous material. The material for the membrane enables the diffusion of biological products of about 6 kDa or less after the manufacturing step. Alternatively, the material for the membrane enables the diffusion of biological products of about 2 kDa or less, about 4 kDa or less, about 6 kDa or less, about 8 kDa or less, about 10 kDa or less, about 12 kDa or less, about 14 kDa or less, about 16 kDa or less, about 18 kDa or less, about 20 kDa or less, about 25 kDa or less, about 30 kDa or less, about 35 kDa or less, about 40 kDa or less, about 45 kDa or less, about 50 kDa or less, about 60 kDa or less, about 70 kDa or less, about 80 kDa or less, about 90 kDa or less, about 100 kDa or less, about 200 kDa or less, about 300 kDa or less, about 400 kDa or less, or about 500 kDa or less after the manufacturing step. The average pore size of the material for the membrane may be about 1 nm or less, about 2 nm or less, about 3 nm or less, about 4 nm or less, about 5 nm or less, about 6 nm or less, about 7 nm or less, about 8 nm or less, about 9 nm or less, about 10 nm or less, about 12 nm or less, about 14 nm or less, about 16 nm or less, about 18 nm or less, or about 20 nm or less after the manufacturing step.

[0045] The manufacturing steps of forming, joining, and cutting the membrane can be performed using one or more devices such as a membrane forming or fusing device. The device can be formed by a 3D printing process, microfabrication, or other machining techniques. In some cases, the device may be in a modular form. In some cases, the shape of the device may affect the dimensions and / or shape of the channels. The device may include a positive and / or negative mold for the membrane (e.g., the first membrane described above). The device may be formed of metal.

[0046] The device may include a platform for the membrane. In some cases, the platform may include a mold for the cell containment device (e.g., a negative mold). In some cases, the platform may include a plate. The platform may further include holes for cutting or molding the membrane. The cutting may affect the diameter (or lumen) of the channels to be formed in the cell containment device. In some cases, the platform may be disposed at a predetermined height and offset from different surfaces. The different surfaces may be surfaces configured such that the membrane is pushed down onto them using the devices described herein. This offset height can determine the depth of the channels in the membrane (e.g., the first membrane).

[0047] The device may include various tools. In some cases, the tool may include a tip or tips for deforming and / or fusing the membrane. The tool can deform the first membrane by pushing down a portion of the first membrane. The tool can include a substantially flat surface configured to be parallel to the first membrane and one or more protrusions on the surface that can deform or push down 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.

[0048] A machine for manufacturing a cell containment device may be configured to support a membrane (e.g., the first membrane described above). The platform may include holes within or below the support, and a second membrane may be disposed. A tool including a plurality of protrusions may be configured to couple with the platform. In some cases, the tool may push down the support platform. Optionally, the protrusions of the tool may be configured to fit into the holes of the platform. In some cases, the first membrane may be disposed on the platform (e.g., above its holes). The tool may push down the first membrane and may form a deformed membrane by pushing a portion of the membrane through the holes. In some cases, the deformed membrane may be fused with the second membrane using the correct pressure and / or temperature described herein.

[0049] A chamber of a molding machine that holds various devices (e.g., the platform and tool described above) and membranes for manufacturing a cell containment device. The chamber may be sealed. The chamber may be configured to hold the platform and / or the tool. The chamber may provide a predetermined or desired temperature necessary for membrane fusion. The molding machine may have a sealed chamber with a gas inlet and / or vent holes. The molding machine may be configured to hold the membrane. For example, the molding machine may include a platform. A flat first membrane may be disposed within the sealed chamber of the molding machine. While the vent holes are closed, nitrogen gas or other gas may be introduced into the sealed chamber through the gas inlet continuously or simultaneously with heating the chamber to a predetermined temperature to reach a predetermined pressure. The membrane may be deformed using the tool, and a deformed first membrane may be generated. In some cases, after the deformation step is completed, the vent holes may be opened to vent the sealed chamber.

[0050] As described above, the first membrane may be deformed using a tool. 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 the location where the tip contacts the membrane. The tip may be in the shape of a cylinder, a cone, or a tapered cylinder or other shapes. The tip may have a contact area at the free end. The contact area of the tip can contact the membrane. The area of the tip is about 0.5 mm 2 or less, about 0.6 mm 2 or less, about 0.8 mm 2 or less, about 1.0 mm 2 or less, about 1.2 mm 2 or less, about 1.4 mm 2 or less, about 1.6 mm 2 or less, about 1.8 mm 2 or less, about 2.0 mm 2 or less, about 2.2 mm 2 or less, about 2.4 mm 2 or less, about 2.6 mm 2 or less, about 2.8 mm 2 or less, about 3.0 mm 2 or less, about 4.0 mm 2 or less, or about 5.0 mm 2 or less may be sufficient. The area of the tip is about 0.2 mm 2 or more, about 0.3 mm 2 or more, about 0.4 mm 2 or more, about 0.5 mm 2 or more, about 0.6 mm 2 or more, about 0.7 mm 2 or more, about 0.8 mm 2 or more, about 0.9 mm 2 or more, about 1.0 mm 2 or more, about 1.2 mm 2 or more, about 1.4 mm 2 or more, about 1.6 mm 2 or more, about 1.8 mm 2 or more, about 2.0 mm 2 or more, about 2.2 mm 2 or more, about 2.4 mm 2 or more, about 2.6 mm 2 or more, about 2.8 mm 2 or more, about 3.0 mm2 Above, about 4.0 mm 2 Above, or about 5.0 mm 2 It may be above. The vertical distance that the tip moves after first contacting the membrane may help determine the height of the channel. The vertical distance that the tip moves after first contacting the membrane may be adjusted to achieve a predetermined height of the channel.

[0051] The first membrane can be deformed into the shape of the channel array. The first membrane can be deformed by thermoforming. The first membrane can be deformed by inflation-based thermoforming. Deforming the first membrane may include pushing down a part of the first membrane using a tool. In one example, the first membrane can be placed in a sealed chamber of a molding machine using a mold to form a channel array. The first membrane can be a flat sheet before the deformation step. The molding machine with the first membrane can be heated to a predetermined temperature critical for deformation. While the vent hole is closed, gas can be introduced continuously and / or simultaneously through the gas inlet into the sealed chamber to reach a predetermined critical pressure and generate the deformed first membrane. In some cases, pressure can be applied by pumping in nitrogen gas. The critical pressure can be applied as a positive or negative pressure. After the deformation step is completed, the vent hole can be opened to vent the sealed chamber. Shown in Figure 8 is a close-up of the pattern of the membrane deforming from a flat configuration to a formed configuration with channels. The flat first membrane 810 is placed on top of the mold 820 in the sealed chamber. After heating the chamber to a predetermined temperature and applying a predetermined pressure to the membrane, the first membrane 811 deforms around the mold to form channels. In some embodiments, thermoforming can form specific internal shapes with set heights and aspect ratios that enable and improve cell adhesion and angiogenesis.

[0052] After the first membrane is deformed, the second membrane can be fused to the first membrane to form a cell containment device. The second membrane may be substantially flat. The second membrane may be disposed on one side of the deformed first membrane. The two membranes can be heated to a sea temperature for fusion. Once a predetermined temperature and / or pressure for fusion is reached, the two membranes can be compressed and / or fused together. The compression of the two membranes can be performed at selected locations across the first membrane. Compression can be facilitated by a mold or plate having the shape and spacing characteristics of the channel array. Heating can be performed in an oven. Alternatively, heating can be performed by a tool with a heating element. The fused first and second membranes can form a fusion at the interface to form compartments. The compartments may be interconnected, resulting in a device that includes a single continuous open space having a volume. The compartments may be enclosed between the base and the top surface of the device, and the compartments may contain cells. FIG. 9 shows the fusion between the formed first membrane 1211 and the flat second membrane 1220, and the resulting device 1250 with the fused first and second membranes.

[0053] The structures of the first and second membranes may be varied by manufacturing process steps. FIG. 10 shows a cross-sectional view of the cell containment device along various portions of the cell containment device. The ultra-fine structure of the PVDF membrane mainly varies from nodules to mainly elongated fibrils (where the material is deformed or fused). The first membrane or the second membrane may include a plurality of nodules interconnected by a plurality of fibrils. During the deformation or fusion step, a part of the nodule structure may change to elongated fibrils. The level of heat or pressure to which the membrane is exposed may affect the number of nodule structures. Generally, in regions of the membrane that have not been deformed or fused (e.g., the upper surface or top of the channel), there may be many observed nodules and few fibrils. Generally, in regions of the membrane that have undergone deformation or fusion (e.g., the bottom of the channel or the middle channel), there are more observed fibrils than in membranes with less deformation or fusion received. Generally, in regions of the membrane that have undergone deformation or fusion (e.g., the bottom of the channel or the middle channel), there are fewer observed nodules than in membranes with less deformation or fusion received. The nodules may be reservoirs of material that can be stretched into fibrils under heat and / or pressure in the deformation or fusion step. Without material migrating from nodules to fibrils within the structure, the membrane may crack during the deformation or fusion step. It is important to perform deformation and fusion without causing cracks in the membrane. The number of nodule and fibril structures and changes in the number of nodule and fibril structures may be specific to the material of the membrane.

[0054] The first and second membranes can be fused without the use of additional adhesives. The first and second membranes can self-seal without the use of an adhesive at a near-sea temperature for fusion. High seal integrity can be obtained by the fusion between the first and second membranes. It may be difficult to visualize the seam between the fused first and second membranes, and as shown in FIG. 11, at the ultrastructural level, the fused portion of the first and second membranes can appear as one continuous membrane by a scanning electron microscope. High seal integrity may allow the cell containment device to be filled at higher pressures. Optionally, an adhesive can be placed between the first and second membranes prior to fusion. In some cases, the adhesive can be pressure-sensitive and / or temperature-sensitive. FIG. 38 shows a tool for sealing the periphery of the device. FIG. 39 shows a scanning electron micrograph of the peripheral edge of the device sealed at 345° C. for 0.5 seconds. It is difficult to identify the seam between the membranes at the ultrastructural level.

[0055] The range of critical pressure and critical temperature may vary for the material used as the membrane. For example, in the case of ePTFE, there may be a desired temperature required for the membrane to retain the deformed shape. In some cases, the desired temperature may be the temperature at which the material is sintered. In some cases, the range of critical pressure and / or critical temperature may vary for each manufacturing step. For the deformation step, the critical pressure may be less than 10 psi, less than 20 psi, less than 30 psi, less than 40 psi, less than 50 psi, less than 60 psi, less than 70 psi, less than 80 psi, less than 90 psi, less than 100 psi, less than 110 psi, less than 120 psi, less than 130 psi, less than 140 psi, less than 150 psi, less than 160 psi, less than 170 psi, less than 180 psi, less than 190 psi, or less than 200 psi. For the deformation step, the critical temperature may be less than 100 °C, less than 110 °C, less than 120 °C, less than 130 °C, less than 140 °C, less than 150 °C, less than 160 °C, less than 170 °C, less than 180 °C, less than 190 °C, or less than 200 °C. For the deformation step, the critical temperature may be less than 210 °C, less than 220 °C, less than 230 °C, less than 240 °C, less than 250 °C, less than 260 °C, less than 270 °C, less than 280 °C, less than 290 °C, less than 300 °C, less than 310 °C, less than 320 °C, less than 330 °C, less than 340 °C, less than 350 °C, less than 360 °C, less than 370 °C, less than 380 °C, less than 390 °C, less than 400 °C, less than 410 °C, less than 420 °C, or less than 430 °C. For the fusion step, the critical temperature may be less than 150 °C, less than 160 °C, less than 170 °C, less than 180 °C, less than 190 °C, less than 200 °C, less than 210 °C, less than 220 °C, less than 230 °C, less than 240 °C, less than 250 °C, less than 260 °C, less than 270 °C, less than 280 °C, less than 290 °C, less than 300 °C, less than 310 °C, less than 320 °C, less than 330 °C, less than 340 °C, less than 350 °C, less than 360 °C, less than 370 °C, less than 380 °C, less than 390 °C, less than 400 °C, less than 410 °C, less than 420 °C, or less than 430 °C.For the fusion step, the critical pressure may be less than 10 psi, less than 20 psi, less than 30 psi, less than 40 psi, less than 50 psi, less than 60 psi, less than 70 psi, less than 80 psi, less than 90 psi, less than 100 psi, less than 110 psi, less than 120 psi, less than 130 psi, less than 140 psi, less than 150 psi, less than 160 psi, less than 170 psi, less than 180 psi, less than 190 psi, or less than 200 psi. It may be necessary to customize the combination of critical pressure and critical temperature for each manufacturing step and the materials used. As the critical pressure and critical temperature range for each material, there may be a range in which the material can be deformed without secondary rearrangement of the crystalline regions of the polymer. In the critical pressure and critical temperature range, the nodes in the thermoelastic material may extend to more fibrillar structures and adapt to new deformed or fused shapes. However, outside the critical pressure and critical temperature range, the material may become crystalline and cracked during deformation or fusion.

[0056] The membrane may have cracks when deformed or fused outside the range of the seashore temperature and pressure against deformation or fusion. Selecting the temperature and pressure outside the range of the seashore temperature and pressure against deformation, in the fusion step, the first membrane may not fuse to the second membrane or the fusion may be insufficient. This crack in the first membrane may not become apparent until the fusion step. Outside the seashore temperature or pressure range, the material of the membrane may have an increased crystallinity. The material for the first membrane with increased crystallinity may have insufficient fusion or no fusion to the material for the second membrane. The relative crystallinity can be measured using differential scanning calorimetry (DSC) to calculate the transition enthalpy of the material. The secondary peak in the DSC heat flow measurement value of the membrane may indicate the rearrangement of the crystal structure of the membrane, indicating that the membrane may not easily fuse with another membrane. This rearrangement may occur without an increase in relative crystallinity. The rearrangement may also occur without a decrease in relative crystallinity. The arrangement of the crystal regions of the membrane is an important factor in fusion, and in the potential rearrangement of the crystal structure, the entanglement of the chains of the crystal regions with another membrane during fusion may not be possible. Another example of the DSC heat flow measurement value of the membrane is shown in FIG. 15. The DSC heat flow measurement value of the membrane deformed at 100 psi and 160 °C maintained a shoulder peak. In comparison, the baseline PVDF membrane had no significant change in its crystal structure and fused with the second membrane. The heat flow measurement value of the membrane deformed at 100 psi and 173 °C and rapidly cooled had different secondary peaks in the first melting endotherm. This indicated that this membrane formed secondary crystal regions and failed to fuse with another membrane.

[0057] In some cases, the first membrane may be deformed using the tip. The tip may move a predetermined vertical distance after first contacting the membrane to reach a predetermined height of the channel. In some cases, the tip may be used to fuse two membranes at the location where the tip causes the membranes to contact each other (the first membrane is already deformed). Fusing may be performed using the tip. The tip presses the first and second membranes against each other for a predetermined time. In some cases, in a single process or step, the tip may be used to deform the first membrane and fuse the first membrane to the second membrane. In some cases, the first and second membranes are vertically offset at a predetermined height. This vertical offset may determine the channel height. Deformation and fusion may be performed in one step using the tip. The tip contacts the first membrane, moves vertically away from the first membrane and towards the second membrane, and presses the first and second membranes against each other for a predetermined time. In some cases, a single tip may be used on the membrane. In FIG. 42, a schematic diagram of a fusion process by spot welding of two membranes using a tip of 2 mm 2 is shown. In other cases, multiple tips may be used on the membrane, for example, simultaneously.

[0058] In some cases, while the membrane(s) is / are stationary, the tip may be moved in the lateral direction (x-y direction) and the vertical direction (z direction). In some cases, while moving the stage holding the membrane(s) in the lateral direction, the tip may be moved only in the vertical direction. In some cases, the tip may move a predetermined lateral distance relative to the membrane surface and then move downward a predetermined vertical distance to enter the membrane and retreat to a neutral vertical position (which may be its previous vertical position). This cycle may be repeated until a predetermined number of channels on the membrane(s) are deformed and / or fused. In some cases, the stage holding the membrane moves laterally to a predetermined location under the tip, the tip moves downward a predetermined vertical distance to enter the membrane, and retreats to a neutral vertical position. This cycle may be repeated until a predetermined number of channels on the membrane(s) are deformed and / or fused. The movement of the tip and / or the stage holding the membrane may be programmed and automated. The tip may move laterally by about 100 μm, about 200 μm, about 300 μm, about 400 μm, about 500 μm, about 600 μm, about 700 μm, about 800 μm, about 900 μm, or about 1000 μm in the x and / or y direction during a cycle. After contacting the membrane, the tip may move vertically by about 100 μm, about 200 μm, about 300 μm, about 400 μm, about 500 μm, about 600 μm, about 700 μm, about 800 μm, about 900 μm, or about 1000 μm.

[0059] When manufacturing a cell-containing device using the tip, there may be critical time, temperature, and pressure ranges for membrane deformation and / or fusion. The time that the tip is in contact with the membrane may be referred to as the tip contact time. In some cases, the tip contact time may be about 0.1 second, about 0.2 second, about 0.3 second, about 0.4 second, about 0.5 second, about 0.6 second, about 0.7 second, about 0.8 second, about 0.9 second, or about 1 second. The tip contact time may be about 2 seconds, about 3 seconds, 4 seconds, 5 seconds, about 6 seconds, about 7 seconds, about 8 seconds, about 9 seconds, about 10 seconds, about 11 seconds, about 12 seconds, about 13 seconds, about 14 seconds, about 15 seconds, about 16 seconds, about 17 seconds, about 18 seconds, about 19 seconds, 20 seconds, about 25 seconds, or about 30 seconds. The tip may be heated to a temperature above the critical temperature for deformation and / or fusion. Substantially as described above, the tip may be heated to a temperature at approximately the critical temperature for deformation and / or fusion. The tip may be heated to about 0 °C, about 1 °C, about 2 °C, about 3 °C, about 4 °C, about 5 °C, about 6 °C, about 7 °C, about 8 °C, about 9 °C, about 10 °C, about 15 °C, about 20 °C above the critical temperature. The tip may apply a pressure of less than 10 psi, less than 20 psi, less than 30 psi, less than 40 psi, less than 50 psi, less than 60 psi, less than 70 psi, less than 80 psi, less than 90 psi, less than 100 psi, less than 110 psi, less than 120 psi, less than 130 psi, less than 140 psi, less than 150 psi, less than 160 psi, less than 170 psi, less than 180 psi, less than 190 psi, or less than 200 psi to the membrane to effect deformation and / or fusion.

[0060] In some cases, the first membrane may or may not be sintered prior to fusion. In some cases, the second membrane may or may not be sintered prior to fusion. In one example, the first membrane may be sintered prior to fusion and the second membrane may not be sintered. The membrane may be sintered at various temperatures for various times. The sintered membrane may have a lower fusion temperature than the same type of non-sintered membrane. In one example, the first membrane may be sintered at 370 °C for 7 minutes. Figure 37 shows the DSC measurement values of ePTFE membranes sintered at 370 °C for 7 minutes and non-sintered at 300 °C. The fusion temperature of the sintered membrane was 320 - 325 °C, which was lower than 340 - 350 °C for the non-sintered ePTFE membrane. Optionally, whether or not to sinter the membrane may be important for fusing the first membrane to the second membrane. In some cases, sintering the membrane may cause the membrane to deform and help maintain the deformed shape. Optionally, fusing a sintered membrane to a non-sintered membrane may result in desired properties (e.g., sealing, device integrity, and / or shape).

[0061] 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 done by punching. The cell containment device may be attached and aligned using an alignment frame, and the excess periphery of the cell containment device may be cut using peripheral punching.

[0062] As shown in FIG. 20, the fusion portion between the first and second membranes can be cut to form the lumen of the channel. The fusion portion between the first membrane and the second membrane may be removed by laser ablation, and as a result, a channel that traverses through the device is formed. FIG. 21 shows a scanning electron micrograph and an image of a cell-containing device with the lumen 2752 of the fusion portion 2754 of the cut by laser ablation and the cell-containing device placed in the frame. FIG. 22 shows a scanning electron micrograph of a cross-section of the cell-containing device after the manufacturing step including cutting through the channel to form the lumen. Cutting can be performed by laser etching or laser ablation. The removed portion can be a part of the region of the fusion portion so as not to impair the seal between the first and second membranes. The removed portion can be about 0%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about 99% of the region of the fusion portion.

[0063] The assembled cell containment device can be obtained as part of a modular system (also referred to herein as a macro device). One or more cell containment devices 2950 are illustrated in FIG. 23. Each can be filled internally with a matrix containing cells that can be assembled onto a frame 2980 to form a macro device. Multiple channel array devices, or cell containment devices with matrices, can be arranged in various configurations on the frame. Each device may have a filling port 2955 for filling the matrix inside the device. The filling port is designed to be small to reduce the possibility of a crack occurring in the device seal. The device may be filled with cells using the filling port. In some cases, the frame may include a flexible backing or a structural support. Optionally, the channel array device can be mounted on a sub-frame 2981 in addition to the frame. The frame may provide flexible support for the channel array device. The frame may prevent unwanted bending of the device. The frame can have one or more flexible mechanisms to prevent buckling of sensitive device areas. The flexible mechanism 2982 can have a break to allow the frame to bend at the break. The flexible mechanism allows the assembly to curve along the tissue at the site of assembly transplantation. The frame can have a handling tab 2956. The handling tab may be used for surgical handling and / or implantation of the assembled device. The frame can have holes to allow for transportation.

[0064] The frame can have a flexible mechanism to prevent buckling of the cell containment device mounted on the frame. The flexible mechanism includes small cuts or cutouts, while a small portion of the frame remains intact. In some cases, the cuts may be at approximately the same location on the upper and lower surfaces of the frame, while the small portion of the frame between the two cuts may remain intact. In other cases, the cuts may be on one surface of the frame, while the small portion of the frame may remain intact. The cuts can be of various shapes, for example, substantially conical, cylindrical, pyramidal, rectangular, or other shapes that remove a part of the frame. Due to the cuts, the frame can bend at various angles (in the range of 0° to 90° in any given direction).

[0065] The macro device can have several configurations and dimensions depending on its application examples. In some cases, the width of the macro device may be at least 3 cm, 4 cm, 5 cm, 6 cm, or 7 cm. In some cases, the length of the macro device may be 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.

[0066] A matrix can be filled into the cell containment device. In some cases, the cell containment device can be filled by pressure. In other cases, the cell containment device can be filled by centrifugation. Figure 24 shows the filling when a PVDF cell containment device with a cell-containing mixture is in the frame (left) and not in the frame (right). In some cases, a filling tube can be connected to the cell containment device to fill the frameless device. In other cases, in a cell containment device placed in a frame, the fluid channel can be connected from the filling hub to the filling port on the device by the frame. Once the device is filled, it can be separated from the filling tube or filling hub and sealed. The frame can also be sealed. The device and / or the frame can be sealed using an adhesive or a UV-curable adhesive. The matrix can include cells or an expression system that produces biological products. The matrix can further include a cell culture medium. The matrix can further include a porous biocompatible material. By leaving only a small area of the cell containment device unsealed for filling, the risk of seal failure can be reduced. After filling the cell containment device with the matrix, the small area can be sealed. Figure 25 shows a cell containment device filled with a matrix with cells visualized by a hematoxylin and eosin (H&E)-stained tissue section. In some cases, the following process can be used to fill the cell containment device (for example, with cells). First, the cells can be suspended in the medium. Second, the cells can be administered into the filling port under pressure. Third, the filling port can be removed. Finally, the filling port opening can be sealed (for example, using a UV-curable adhesive).

[0067] The assembled channel array device or its components can be processed on its surface. The surface can be processed with a material to promote angiogenesis. The coating can be VEGF, or other angiogenesis-promoting factors or substances. The outer surface can be processed with a material that provides anti-fouling properties. The surface of the device can be processed to reduce the likelihood of fibrosis occurring around the device or the likelihood of connective tissue being formed. The material of the device can be selected to reduce the likelihood of fibrosis. The surface of the device can be processed to form physical features or chemically processed to reduce fibrosis. The surface can be processed with a hydrophilic coating. The hydrophilic coating may include a polymer, polyethylene glycol, polyvinyl alcohol, polydopamine, oact. Figure 41 shows an example of a protocol for forming a hydrophilic coating on the surface of a membrane. Prior to the coating treatment, the hydrophilic coating may impart hydrophilicity to the hydrophobic surface. Improving the hydrophilicity of the membrane surface may improve the transport of hydrophilic molecules across the membrane between the internal and external environments of the device. Figure 38 shows an ePTFE cell containment device after hydrophilic coating treatment in water. The bubbles indicate the ability to fill the interior of the device with water and move air out of the interior of the device.

[0068] One or more porous materials can be included in the membrane of the cell containment device. The membrane may include PTFE, ePTFE, PVDF, PCL, PE / PES, PP, PS, PMMA, PLGA, PLLA, or other thermoelastic materials. The materials for the membrane may be synthesized in various ways. The synthesis methods of the porous material may include an expansion method, a solution casting method, an immersion precipitation and phase separation method, an electrospinning method, a method for obtaining an isotropic network, a method for obtaining a columnar network, or other methods. The membrane is a porous material that, after the manufacturing process, can transport materials with a molecular weight of less than about 3000 kDa, less than about 2000 kDa, less than about 1000 kDa, less than about 500 kDa, less than about 400 kDa, less than about 300 kDa, less than about 200 kDa, less than about 100 kDa, less than about 50 kDa, less than about 40 kDa, less than about 30 kDa, less than about 20 kDa, less than about 10 kDa, less than about 6 kDa, less than about 5 kDa, less than about 4 kDa, less than about 3 kDa, less than about 2 kDa, or less than about 1 kDa through this material. The average pore size of the membrane may be about 5 nm, about 10 nm, about 15 nm, about 20 nm, about 30 nm, about 40 nm, about 50 nm, about 60 nm, about 70 nm, about 80 nm, about 90 nm, about 100 nm, about 200 nm, about 300 nm, about 400 nm, about 500 nm, about 600 nm, about 700 nm, about 800 nm, about 900 nm, about 1000 nm, about 1100 nm, about 1200 nm, about 1300 nm, about 1400 nm, about 1500 nm, about 1700 nm, about 2000 nm, or about 2500 nm.

[0069] 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 channels. The pressure during the deformation step can be used to change the depth of the channels. A combination of temperature and pressure during the deformation step can be used to change the depth of the channels. The temperature and pressure during deformation may be specific to the material of the first membrane. The change in channel dimensions can affect the three-dimensional shape of the interconnected pockets within the cell containment device. These manufacturing parameters can be used to adjust the configuration and dimensions of cell containment for the channel array device. Using these parameters, the SA:V ratio of cell containment can be changed. The SA:V ratio of cell containment may improve angiogenesis within and around the device.

[0070] The device can be implanted at various sites within the subject's body. In some cases, the device on the frame can be implanted in the subject. In one example, the device can be placed by preperitoneal or retrorectus implantation. In other examples, the device can be placed by intraperitoneal implantation. In another example, the device can be placed by subcutaneous implantation. In another example, the device can be placed by suprahepatic implantation. Figure 28 shows preperitoneal, intraperitoneal, suprahepatic, and subcutaneous implantation of the cell containment device into rats.

[0071] The device can be fixed at the implantation site in the body. In one example, a tissue adhesive can be used to fix the device. The tissue adhesive can be fibrin, cyanoacrylate, polyethylene glycol, albumin-based adhesive, or polymer-based adhesive. In another example, platelet-rich plasma can be used to fix the device.

[0072] After in vivo transplantation, blood vessels may form around the device and through channels within the device. Figures 31 and 32 show the vasculature around and through the channels of the cell containment device 20 and 90 days after transplantation into the preperitoneal site in rats. Figure 29 shows an H&E stained tissue section of angiogenesis around the cell containment device and vasculature within the channels after in vivo transplantation into rats. Figure 30 shows angiogenesis around low-channel density and high-channel density cell containment devices after in vivo transplantation. The blood vessels may have smooth muscle cells, which are generally found in arteries. The blood vessels may have arterial characteristics. Figures 30 and 31 show that the density of channels within the channel array device can affect the level of angiogenesis. The higher the density of channels, the higher the level of angiogenesis may be. The lower the density of channels, the lower the level of angiogenesis may be. The diameter of the channels within the channel array device can affect the level of angiogenesis and branching of the blood vessels.

[0073] The cell containment device can be designed to achieve various functional goals. One goal of the cell containment device is to provide at least one year of cell viability for the cells within the device after in vivo transplantation. In some embodiments, the device is designed to be able to have at least 4x10 8 cells. The device may be retrieved or explanted from the in vivo transplantation site of the subject to evaluate cell viability and other functional evaluations after transplantation. The cell containment device may be designed to improve mass transport within the device and through the membrane of the device. The cell containment device may be coated and / or transplanted at a site that improves proximity to the host vascular supply. The cell containment device may be designed to stabilize the host / device interface such that flexibility within the device is possible without folding in on itself. The cell containment device may be designed to suppress tissue integration such that it provides stability to the device without including the integrity of the device. The cell containment device may be designed to use materials and coatings that result in favorable non-specific biomaterial reactions.

[0074] Ultra-thin device

[0075] Although this specification mainly describes a cell-containing device including a channel, the cell-containing device does not necessarily have to include a channel. For example, in some cases, another approach to meet various functional goals of the cell-containing device can be achieved using an ultra-thin cell-containing device (also referred to as an ultra-thin device herein). Accordingly, various parameters, characteristics, or descriptions (such as coatings, materials, etc.) described for a given embodiment of a cell-containing device (e.g., one including a channel) can be similarly applied to another embodiment of the cell-containing device (e.g., an ultra-thin device).

[0076] The ultra-thin device may have a thin overall cross-sectional thickness. Alternatively or in addition, the membrane of the ultra-thin device may be thin. The membrane of the ultra-thin device may be a biocompatible polymer or biomaterial, such as ePTFE, PVDF, PEEK, PS, PES, PAN / PVC, nylon, polyurethane, polycarbonate, polyacrylonitrile, glass fiber, polycaprolactone, hydrogel, polyester, polyanhydride, or cellulose. Also, the membrane may be formed from a permanent non-degradable material or, alternatively, a biodegradable material with a controlled degradation profile. A high SA:V ratio may be obtained depending on the dimensions of the ultra-thin device. The high SA:V ratio can enhance the transport of molecules into and out of the device, for example, the transport of nutrients and oxygen into the device for the resident cells within the device, and the transport of insulin or other secreted products out of the device. The modeled insulin diffusion out of an ultra-thin device filled with insulin-producing cells can be 0.4 - 10 ng / cm 2 / 10 minutes. The ultra-thin device may not have channels that run through the thickness of the device as in the case of a channel array device. Figures 57 and 58 show schematic views of an ultra-thin device (black circles) with a cross-sectional thickness of 250 μm filled with cells and a macro device with three ultra-thin devices.

[0077] The ultra-thin device may have a total cross-sectional thickness of about 250 μm. In some embodiments, the total cross-sectional thickness of the ultra-thin device is less than 5000 μm, less than 4000 μm, less than 3000 μm, less than 2000 μm, less than 1000 μm, less than 900 μm, less than 800 μm, less than 700 μm, less than 600 μm, less than 500 μm, less than 400 μm, less than 300 μm, less than 200 μm, less than 100 μm, or less than 50 μm. In some embodiments, the total cross-sectional thickness of the ultra-thin device is 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.

[0078] The ultra-thin device may have a membrane thickness in the range of 2 μm to 25 μm. In some embodiments, the membrane of the ultra-thin device is less than 100 μm, less than 90 μm, less than 80 μm, less than 70 μm, less than 60 μm, less than 50 μm, less than 40 μm, less than 30 μm, less than 20 μm, less than 10 μm, less than 5 μm, or less than 1 μm.

[0079] The ultra-thin device may be designed such that the SA:V ratio is an appropriate value for transporting nutrients and desired products through the device. In some cases, the SA:V ratio may be 80 cm -1 or more. In other cases, the SA:V ratio is about 20 cm -1 or more, about 40 cm -1 or more, about 60 cm -1 or more, about 80 cm -1 or more, about 100 cm -1 or more, about 120 cm -1 or more, about 150 cm -1 or more, or any value in between. The maximum oxygen diffusion distance of the device may be less than 50 μm, less than 100 μm, less than 150 μm, less than 200 μm, less than 250 μm, less than 300 μm, less than 350 μm, less than 400 μm, less than 450 μm, or less than 500 μm.

[0080] The characteristics of the membrane of the ultrathin device may be selected to enhance its function. One such characteristic may be the flux selectivity of the membrane. Various membrane characteristics (e.g., microstructure, twist, pore size, porosity, and / or thickness) may contribute to flux selectivity. Flux selectivity may affect the molecules that can pass through the membrane. Since the microstructure affects the flux selectivity of the membrane, the treatment of the membrane may affect its microstructure and its flux selectivity.

[0081] The characteristics of the membrane of the ultrathin device may be selected to improve its ability for immune protection. For example, if the flux selectivity of the membrane is high, a large amount of antibodies and complement proteins can be prevented from moving across the membrane. Also, if the flux selectivity of the membrane is high, the diffusion of nutrients through the membrane to the inside of the cell containment device can be reduced. When the flux selectivity of the membrane is intermediate, the flux characteristics for cell survival are favorable, the release of antigens from dead cells is reduced, and the movement of antibodies across the membrane can be prevented at a certain level. If the flux selectivity of the membrane is low, it may be possible for the exchange of nutrients through the membrane to be promoted by high flux, the transport of cells across the membrane can be restricted, and the transport of molecules smaller than cells may not occur. Also, when the flux selectivity of the membrane is intermediate or low, potential exchange of antibodies may be possible, and the mechanical properties may be lower compared to the case where the flux selectivity of the membrane is high.

[0082] For ultra-thin devices, various types of membranes can be used. Figure 47 shows scanning electron micrographs of ePTFE membranes with different porosity thicknesses that can be used for ultra-thin devices. Table 1 shows various membrane characteristics, such as their flux, thickness, diffusion selectivity, and insulin flux. Diffusion selectivity is defined as the ratio of antibody flux to insulin flux. The lower the ratio, the higher the selectivity of the flux. This is because insulin is a much smaller molecule than an antibody. Membrane C has a lower diffusion selectivity than membranes A and B and is highly selective in allowing larger molecules (e.g., antibodies) to be transported across the membrane.

Table 1

[0083] As shown in FIG. 48, a high-flux ultra-thin device containing a selective permeation membrane and filled with rat pancreatic islet cells was able to produce and secrete insulin in response to a 20 mM glucose stimulus. The measurement was performed by the flux of C-peptide to the outside of the ultra-thin device. C-peptide (also known as the connecting peptide) is a polypeptide that is cleaved from proinsulin to form insulin molecules. Since C-peptide is present in an equimolar amount to insulin, the level of C-peptide indicates the level of insulin produced and secreted. Using islets taken from Sprague-Dawley rats, the insulin transport kinetics after encapsulation into the ultra-thin device were evaluated. The ultra-thin device filled with encapsulated rat pancreatic islet cells showed that the release of insulin from the ultra-thin device was delayed by about 5 - 10 minutes compared to the ultra-thin device filled with free rat islets. The measurement was performed as dynamic GSIS (glucose-stimulated insulin secretion) over time. Encapsulated pancreatic islet cells were able to produce and release insulin in response to a glucose stimulus. Encapsulation did not prevent the ability of the encapsulated cells to receive a glucose stimulus and respond to the glucose stimulus by producing and secreting insulin. The ultra-thin device filled with pancreatic islet cells has diffusion kinetics that allow for the conversion of biphasic insulin secretion and blockade depending on the glucose concentration in its environment. GSIS occurs when cells secrete insulin under exposure to glucose. The amount of insulin secreted may be proportional to the level of glucose exposure. During GSIS, insulin secretion may decrease, stop, start, and increase according to the level of glucose exposure. Some embodiments further include releasing insulin from the cells in the cell containment device in an amount sufficient to lower the blood glucose level of a subject. In some embodiments, the release of insulin stops when the blood glucose level of the subject has dropped to a normal level. In some embodiments, the release of insulin resumes when the cells in the cell containment device are re-exposed to the high blood glucose level of the subject.

[0084] The selection of the membrane may affect the flux dynamics of insulin from the ultrathin device and the responsiveness of the ultrathin device to glucose stimulation. Figure 49 shows the static C-peptide release in an ultrathin device filled with encapsulated pancreatic islet cells, including a high-flux membrane or a selective permeation membrane, before high glucose stimulation (LG or low glucose), during high glucose stimulation (HG), and after high glucose stimulation (LG). Encapsulated pancreatic islet cells in all three types of ultrathin membranes responded to glucose stimulation. The ultrathin device with a high-flux membrane reached approximately 1.7x10 5 pM of C-peptide per device under HG conditions, while the ultrathin device with a selective permeation membrane reached approximately 5x10 4 pM of C-peptide per device under HG conditions. In the case of the ultrathin device, a delay occurred in insulin release in response to the HG condition. The delay in insulin release may be due to retaining insulin within the ultrathin device, indicating that some membranes may have adhesiveness to insulin. Figure 50 shows the dynamic GSIS of the ultrathin device with an AS1 membrane. Since AS1 is a high-flux membrane, this results in a high-flux ultrathin device that responds to a 20 mM high glucose stimulation by generating and secreting insulin and turns off insulin generation and secretion when the high glucose stimulation is removed.

[0085] Ultra-thin devices and cell-containing devices can be prepared by many processes. These processes can include membrane fabrication, membrane coating, device assembly, and aseptic cell filling. The membrane can be fabricated and processed to achieve the targeted properties. The targeted properties may include transport flux properties, mechanical properties, porosity, pore size, thickness, microstructure, or twist. The fabrication process may include membrane stretching or sintering. The membrane can be further processed using a coating process to impart various desired properties. As described above, these coatings may include hydrophilic polymers, VEGF, or other molecules that promote angiogenesis or protein transport. Then, the membrane can be fabricated and assembled into a device. The assembly of the ultra-thin device can be performed using robotic assembly that can be automated or semi-automated. A frame for the device can be fabricated. Optionally, one or more ultra-thin devices can be assembled on the frame as a macro device. Cells can be aseptically filled into the ultra-thin device. The aseptic cell filling may be performed by a pressure filling process, centrifugation, gravity filling, open filling, or any combination thereof.

[0086] The membrane may be sintered before being fabricated into a cell-containing device. The sintering process of the membrane may be used to change the porosity and flux properties of the membrane. By sintering, the porosity of the membrane can increase while maintaining its pore structure. By sintering, the mechanical stability and insulin flux of the membrane can be improved. Figure 51 shows scanning electron micrographs of non-sintered and sintered membranes, showing the change in the microstructure of the membrane. There are fusions and coalescences in the nodules and fibrils of the sintered membrane.

[0087] The sintering process is highly consistent and can reduce lot-to-lot variation. Shown in Table 2 are the melting point temperatures of the sintered membranes (in the range of approximately 326°C to 333°C), which are different from those of the non-sintered membranes (approximately 345°C) (measured by DSC). As a result of the sintering process, the lot-to-lot variation was 0.76%, indicating the consistency of the sintering process. The porosity of the membrane can be changed using membrane sintering, and then it can be used to adjust the porosity and flux characteristics of the cell containment device. The consistency in the sintering process may provide an attractive option that affects membrane porosity and properties in the mass production of cell containment devices.

Table 2

[0088] In some embodiments, the cell containment device assembled on the frame may be made using a non-sintered membrane and then passed through post-curing to reduce the porosity on the frame. This manufacturing method of the assembly using a non-sintered membrane and post-curing may reduce the steps, time, and / or cost in the overall manufacturing process.

[0089] In some embodiments, the cell containment device may include at least one sintered membrane and at least one non-sintered membrane. Such asymmetric sintering of the membranes of the cell containment device may result in a device with a controlled geometric shape. By using different types of membranes within one cell containment device, curvature may be induced within the device due to their different mechanical properties. The first membrane may be more flexible or ductile than the second membrane. In some embodiments, the different types of membranes within one device may be sintered membranes and non-sintered membranes.

[0090] By coating the membrane, another approach to adjusting the flux characteristics of the cell containment device is obtained. The membrane may be coated with a hydrophilic coating before being fabricated into the cell containment device. The hydrophilic coating enables the membrane to be wetted, allows for ultrafiltration to be utilized to fill the device with cells, allows for diffusion, and may result in a biocompatible neutral charge surface. Figure 52 shows a comparison of the hydrophilic coating process for a membrane near a target insulin flux of 1x10 -6 mol / m 2 / s. The hydrophilic coating process resulted in within-lot variation of approximately 9% and between-lot variation of approximately 8%.

[0091] In some embodiments, a hydrophobic membrane can be coated with a hydrophilic coating. The hydrophilic coating can be biocompatible and can improve the diffusion of insulin and other molecules. In some embodiments, with an uncoated hydrophobic membrane, the diffusion of insulin and other molecules may not be possible. In some embodiments, a nano-thin coating process may result in an appropriate level of permeability and insulin diffusion for the membrane and the cell containment device. In some embodiments, the semi-permeability of the membrane is configured to protect the cells from immune attack. In some embodiments, the semi-permeability of the membrane is configured to protect the cells from immune attack in the absence of immunosuppressive therapy.

[0092] Since the ePTFE material is hydrophobic, a hydrophilic polymer can be polymerized around the ePTFE microstructure to enable wetting of the membrane and reduce hydrodynamic resistance. As a result, ultrafiltration during the cell filling process can be facilitated, and it can be possible to introduce cells into the device using low pressure. It is also possible to form a neutral hydrophilic surface to minimize the adsorption or attachment of host proteins and cells.

[0093] The sintered membrane within the frame may be placed in 100% ethanol for 5 minutes and then immersed in 30% ethanol for approximately 5 minutes. The membrane can then be immersed in a coating solution consisting of 9 g APS, 27 mL of HPA, and 18 mL of TEGDA in 30% ethanol at room temperature for approximately 5 minutes. The polymerization reaction can be controlled using LabView software and carried out at 70 °C with an increase rate of 3 °C / min from room temperature. The coated membrane within the frame can be removed from the coating solution, transferred to boiling 100% ethanol to remove unreacted monomers, and then immersed with several changes of excess distilled water. Finally, the coated membrane is dried in the chamber using a continuous nitrogen stream.

[0094] Figure 53 shows membranes coated by three different coating processes V1, V2, and V3 stained with H&E dye as an indication of membrane hydrophilicity. The membrane subjected to the V1 coating process appeared highly stained pink and seemed to be overcoated. The membrane subjected to the V2 coating process had variable levels of H&E dye staining, with its edges being stronger than the center and seemed to have a gradient coating. The membrane subjected to the V3 coating process was coated by a nano-thin coating process and seemed to be uniformly coated throughout its cross-sectional thickness. Figure 54 shows the water permeability of the membrane, the membrane subjected to the V1 coating process, and the membrane subjected to the V3 coating process. The V3-coated membrane showed a high permeability of about 1.8x10 -14 m 2 while the membrane and the V1-coated membrane showed low water permeabilities of less than 1x10 -15 m 2 .

[0095] In some embodiments, the water permeability of the coated membrane in the ultra-thin device is at least 1x10 -16 m 2 , 1x10-15 m 2 、 1 x 10 -14 m 2 、 or 1 x 10 -13 m 2 may also be. In some embodiments, the first and second membranes of the ultrathin device may have the same water permeability. In some embodiments, the first and second membranes of the ultrathin device may have different water permeabilities. In some embodiments, different coating processes may be used on the first and second membranes to achieve different water permeabilities. In some embodiments, the semipermeability of the first membrane, the second membrane, or both is configured to protect cells from immune attack. In some embodiments, the semipermeability of the first membrane, the second membrane, or both is configured to protect cells from immune attack in the absence of immunosuppressive therapy.

[0096] The coating process may be designed and scaled up to coat multiple membranes or multiple cell containment devices at once. In some embodiments, the coating process may be scaled up to coat 40 human cell containment devices at once. Figure 55 shows a setup for scaling up the coating process with dry and wet coated membranes. The high light transmission observed by transmission illumination of the wet coated membrane demonstrates the wettability of the coated membrane, which functions as an indication of the hydrophilicity and permeability of the coated membrane.

[0097] The frame for the cell containment device may include various materials. In some embodiments, the frame may be a biocompatible material. As described above, the frame may hold one cell containment device or multiple cell containment devices. The mechanical properties of the frame may be similar to the host biological tissue surrounding the device after in vivo implantation. One indicator of the mechanical properties of a material is its Young's modulus. FIG. 56 shows the Young's modulus of various biological and synthetic materials, as well as the ranges of the target Young's modulus (composite Young's modulus) for the candidate membrane, candidate frame material, and the entire device. The target Young's modulus for the membrane may be in the range of 10 6 ~10 9 Pa. The target Young's modulus for the frame material may be in the range of 10 8 ~10 9 Pa. The target Young's modulus for the device composite (also referred to as the macro device or the device with the frame) may be in the range of 10 7 ~10 9 Pa (the middle of the range for the device alone and the range for the frame alone). In some embodiments, the Young's modulus of the membrane may be at least 10 5 Pa, 10 6 Pa, 10 7 Pa, 10 8 Pa, or 10 9 Pa. In some embodiments, the Young's modulus of the frame may be at least 10 7 Pa, 10 8 Pa, or 10 9 Pa. In some embodiments, the Young's modulus of the device composite may be at least 10 7 Pa, 10 8 Pa, or 10 9 Pa.

[0098] In some embodiments, the frame may include polyetheretherketone (PEEK). FIG. 57 shows a simulation of the mechanical properties of a 300 μm thick PEEK frame for two cell containment devices under a force of 150 mN. As shown by the simulation, the PEEK frame has a maximum stress of about 88 MPa (below the yield stress of about 103 MPa), a maximum strain of about 0.014, and a maximum displacement of about 4.711 mm.

[0099] FIG. 58 shows an example of a PEEK frame holding a cell containment device with a fused dot at the center of the device. The frame may be microfabricated or machined using an appropriate method to achieve its target dimensions. FIG. 59 shows a single frame module including a single cell containment device with a dot fused at the center on a maximally filled frame. The filled device with a dot fused at the center on the frame indicates that the lateral expansion of the membrane is limited.

[0100] FIGS. 89A and B show various configurations of a macro device with multiple devices held by a macro device frame. In one design, the macro device frame is flexible and can hold multiple devices. In some embodiments, the macro device frame can be a flexible integrated frame for holding multiple devices and can have a porous structure surrounding individual cell containment devices.

[0101] As shown in FIG. 59, the cell-containing device may be filled with cells to the maximum extent. FIG. 60 shows the cell viability for at least 10 days when stored under standard conditions of 20% oxygen and 37° C. in the cell-containing device filled to the maximum extent. This is evident from the presence of cells in the H&E-stained tissue sections. The cell-containing device filled with cells may be stored under various conditions to extend the viability of the cells in the cell-containing device before transplantation. The cell-containing device filled with cells may be stored at various temperatures, such as 4° C., 23° C., or 37° C. In some embodiments, the storage temperature of the cell-containing device filled with cells may be 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-containing device filled with cells may be stored under hypoxic, normoxic, or hyperoxic conditions.

[0102] The two membranes of the cell-containing device may be fused along their surfaces into separate dots. Various configurations of the fused dot shape, diameter or distance, and density (e.g., the distance between centers) of the device may exist. The dots may be circular, rectangular, triangular, linear, or other shapes. The dots may have various cross-sectional distances or diameters. In some embodiments, the dot diameter may 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.

[0103] The device may have one fused dot or multiple fused dots. The dots may be regularly spaced. The dots may be regularly spaced within a matrix array. The dots may be irregularly or randomly spaced. In some embodiments, the dots are spaced such that the distance between centers may be 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. In some embodiments, the dots may be arranged to overlap each other.

[0104] The total surface area of the dots may cover a portion of the surface area of the membrane of the cell-containing device. The surface area of the membrane of the cell-containing device covered by the dots may be a portion that does not interfere with its ability to maintain cell viability, cell function, and the release of molecules from inside the device. In some embodiments, the dots may cover less than 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% of the surface area of the membrane.

[0105] The dots may provide adhesion limitations such that the membrane of the device does not bend away from each other. The dot density and diameter can be selected to limit the deformation, bending, or expansion of the device's membrane away from each other during filling of the device. The dot diameter and density can affect the acceptable fill volume by forming a series of counters. For devices of the same dimensions, a device with a shorter dot pitch may have a higher dot density than a device with a longer dot pitch. For devices having the same dot diameter and device dimensions, a device with a shorter dot pitch can have a smaller internal volume available for filling than a device with a longer dot pitch.

[0106] The dimensions of the dots may be adjusted to control the volume and SA:V ratio within the cell containment device. In some embodiments, the surface area of the dots may be increased to decrease the volume within the cell containment device. As a result, an increase in the surface area of the channels may increase the SA:V ratio for the cell containment device. In some embodiments, the surface area of the dots may be decreased to increase the volume within the cell containment device. A decrease in the surface area of the dots may decrease the SA:V ratio for the cell containment device.

[0107] The dots may be formed in various patterns. The dots may be formed in a pattern that maintains the ability to uniformly fill the device across the entire device. The dot pattern can be designed to adjust the volume and / or amount of cells that can be filled into the device. The dot pattern can be designed to prevent the membrane of the device from bulging or bending during filling of the device with cells. In some embodiments, the device may have one dot. In some embodiments, the device may have multiple dots. In some embodiments, the device may 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 dots.

[0108] The thickness of the device during filling can further be controlled by applying an external restraint to the outer surface of the device. The external restraint can be placed on the outer surface of the membrane of the cell containment device to cover most or all of the surface of the membrane. The external restraint may physically limit the membrane of the device from deforming away from each other during filling. The external restraint can be made porous to assist in the release of air that has moved out of the device when the device is filled. The external restraint can be made further adjustable by using spacers with a target spacing between the porous restraints. The spacers may correspond to the total thickness of the device before filling.

[0109] The dots may be formed by various processes so as to fuse the membranes at separate dots. The dots may be formed by a fusion process using the spot welding process described herein. The dots may be formed by placing an adhesive at a desired location relative to the dots on the first membrane and then bringing the first membrane into contact with the second membrane. The adhesive dots can be stacked in multiple layers. In some embodiments, the adhesive dots are stacked in two layers. In some embodiments, the adhesive may be placed on the membrane by an automated or semi-automated process. In some embodiments, an automatic dispenser may be programmed to dispense a specific volume or weight of adhesive at a specific location. In some embodiments, information regarding the dimensions of the membrane and dot pattern may be provided to the dispenser to guide the dispensing of the adhesive onto the membrane. The dots may be formed by a combination of adhesive placement and the spot welding process.

[0110] Many adhesives may be suitable for forming the fused dots. The adhesive may be a cyanoacrylate, urethane acrylate, UV curable epoxy, thermosetting epoxy, or two-part epoxy (one element may be implanted (monomer) within the membrane and the other may be applied in a dissolved state around it (crosslinking agent)). Alternatively or in combination, a solvent may be used to partially solubilize the membrane bond to form a bond.

[0111] The dot diameter, density, and the thickness of the device during filling can be adjusted using the properties of the adhesive and the pattern of adhesive placement. Adhesiveness can affect the rate of adhesive penetration into the membrane. The effective diameter of the dot may be set by the initial application. The penetration rate can be made a function of the viscosity of the adhesive. Depending on the chemical nature of the adhesive, the penetration rate may be affected by the charge and degree of hydrophilicity of the membrane. The coating, membrane, and adhesiveness all shape the density of the dot pattern. Alternatively or in addition, the dot pattern can be controlled by a picosecond adhesive dispenser that adjusts these parameters over time, and the dispensing by the dispenser can be made non-linear dispensing.

[0112] In some embodiments, the adhesive has a viscosity of about 200 cP to 450 cP. In some embodiments, the adhesive has a viscosity of 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.

[0113] The cell containment device can be scaled to various sizes while maintaining various parameters constant. For example, the device can be scaled such that a microenvironment equivalent to that of a device for human transplantation is provided within a device for mouse transplantation. Various parameters important for the function of the device can be maintained constant. These parameters can be the diffusion distance of a molecule of interest such as insulin or the SA:V ratio. Since the size of the device may affect the mechanical properties, the device may be designed such that the flexibility of the device is maintained without overlapping at the target transplantation site. Since the filling process is also scaled up, the scaled-up device can be designed to be compatible with filling of the device at a higher throughput. In the design of the scaled device, updated fluid channels may be incorporated to improve the final seal of the device after filling. As shown in FIGS. 74A and B, in the design of the scaled device, an integrated macro device frame for a plurality of devices may be incorporated instead of a basic flexible macro device frame.

[0114] Provided herein is an ePTFE cell containment device comprising a first membrane and a second membrane facing 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 opposing second surface having a surface area. The first membrane and the second membrane may form an enclosed compartment for providing a volume for containing cells within the ePTFE device.

[0115] A typical PVDF channel array device may attenuate under UV light.

[0116] Although ePTFE transfer may be enabled by a typical frameless ePTFE channel array device, the device may be distorted in vivo depending on the situation. In a typical frameless ePTFE channel array device, a stable frame and filled tubes with straight and angled sections are each used. Improved fluid flow may be obtained with the angled filled tubes. Additionally, the angled filled tubes may increase the dead space within the filled tubes attached to the membrane. Finally, the angled filled tubes may reduce the need for laser vision during manufacturing. As shown, the angled filled tubes are oblique to the symmetric bisecting plane of the membrane, and the straight filled tubes are in the same plane or parallel to the symmetric bisecting plane of the membrane.

[0117] This specification further provides a method of manufacturing a cell containment device. The method may include providing a first membrane, forming a plurality of channels within the first membrane, and fusing a second membrane to the first membrane. The first membrane may include a first surface and a second surface. The second surface may face the first surface. A plurality of channels may be formed within the first surface of the first membrane. The second membrane may be fused to the second surface of the first membrane. A compartment may be formed by fusing the second surface of the first membrane and the second membrane. The compartment may be configured to contain cells between the second surface of the first membrane and the second membrane.

[0118] The formation of a plurality of channels within the first surface of the first membrane is performed by molds 1021, 1022. The molds may include a positive mold 1021 (seen in FIG. 86A) or a negative mold 1022 (seen in FIG. 86A). The positive mold 1021 may contact the second surface of the first membrane, while the negative mold 1022 may contact the first surface of the membrane. By using the negative mold 1022, 3D thermoforming of the membrane may be improved, and a membrane with more optimal thickness and porosity may be produced.

[0119] It may be possible to insert cells into the device via an external filling tube by means of the upper part of a typical channel array device. A membrane is included in the lower part of a typical channel array device. The illustrated external filling tube may include an angled filling tube. The filling tube is oblique from the symmetric bisecting plane of the membrane. The upper part of a typical channel array device may further function as a mechanical frame for support. A typical channel array device may have an internal volume of about 24 μl, a channel quality of about 50, a footprint of about 1 x 0.5 cm, channels of 1 mm, and a channel-to-channel (C-C) distance of about 1 mm. As seen in FIG. 87, a typical channel array device includes a frame. The insertion tube 1040 may be manually attached onto the frame. A typical channel array device shown in FIG. 87 may include a serrated channel array device.

[0120] The pocket frame may enable the incorporation of fluid channels within the channel array device. Further, the pocket frame is easier to manufacture without the need to manually add insertion tubes. Multiple such channel array devices may be formed at once. Multiple simultaneously formed channel array devices may be easily filled, separated, and sealed, enabling the manufacture of self - contained and / or mechanically actuated devices. The pocket frame within the channel array device may be formed by selective laser sintering (SLS), injection molding, solution casting, machining, or any combination thereof. Such processes may enable a high enough control over the geometry and resolution of the pocket frame.

[0121] Further, the pocket frame may be configured to provide torsional resistance, stiffness - flexibility, or both, to the channel array device through its hoop strength. Such resistance may be obtained with a pocket frame having variable thickness, width, cross - sectional shape, or any combination thereof. Further, the pocket frame sealed between the first and second membranes can be configured to be airtight to prevent contamination.

[0122] Finally, as shown in FIGS. 106a and 106b respectively, the pocket frame 1051 within the channel array device 1050 can make the mass flow rate larger and more uniform than the channel array device 1050 with a filling port.

[0123] As shown in FIGS. 107A - B, the geometric design parameters 1070 of the channel array device are optimized to enhance effectiveness. The geometric shape of the channels within the channel array device 1070 may be defined by the cell chamber height (A), the fusion region (B), the opening diameter (C), the channel diameter (D), and the channel spacing (E).

[0124] The cell chamber height (A) may be measured as the maximum normal distance between the inner surfaces of the first membrane 1071 and the second membrane 1072. The cell chamber height (A) may be measured as the average value of the maximum normal distances between the inner surfaces of the first membrane 1071 and the second membrane 1072 for all the channels 1073 in the channel array device 1070. In some embodiments, the cell chamber height (A) is at least about 300 μm. As shown in Table 3 below, the cell chamber height (A) may be optimized to vary the diffusion flux, foreign body reaction, angiogenesis, non-traumatic cell filling, and volume / footprint of the device.

[0125] The fusion region (B) may be measured as the total surface area where the first membrane 1071 and the second membrane 1072 are fused. Alternatively, the fusion region (B) may be measured or correlated as the surface area of the upper surface of the first membrane 1071 substantially parallel to the second membrane 1072. As shown in Table 3 below, the fusion region (B) may be optimized to vary the seal integrity, foreign body reaction, and angiogenesis of the device.

[0126] In some embodiments, the device 1070 further includes an opening 1074 passing through the first membrane 1071 and the second membrane 1072 in the channel 1073. The opening 1074 may have an opening diameter (C). The opening diameter (C) may be measured as the average, maximum, or minimum inner diameter of the opening 1074. The opening diameter (C) may be measured as the average, maximum, or minimum opening diameter (C) of the plurality of openings 1074 in the plurality of channels 1073 in the device 1070. In some embodiments, the opening 1074 has a maximum concentricity with respect to the channel 1073 and is 25% of the channel diameter (D). As shown in Table 3 below, the opening diameter (C) may be optimized to vary the seal integrity, foreign body reaction, and angiogenesis of the device.

[0127] In some embodiments, the channel diameter (D) may be measured as the maximum, minimum, or average inner diameter of channel 1073. In some embodiments, the channel diameter (D) may be measured as the maximum, minimum, or average normal inner diameter of channel 1073. In some embodiments, the channel diameter (D) is measured at the narrowest point within the channel. In some embodiments, the channel has an average diameter of from about 400 μm to about 3,000 μm. As shown in Table 3 below, the channel diameter (D) may be optimized to vary the diffusion flux, foreign body reaction, angiogenesis, non-traumatic cell seeding, and volume / footprint of the device.

[0128] The channel spacing (E) may be measured as the maximum, minimum, or average distance between the inner surfaces of two adjacent channels 1073. The channel spacing (E) may be measured as the maximum, minimum, or average normal distance between the inner surfaces of two adjacent channels 1073. The channel spacing (E) may be measured as the average value of the maximum, minimum, or average distances between the inner surfaces of two adjacent channels 1073 for each of a plurality of channels 1073. In some embodiments, the centers of each channel are separated by a distance of from about 75 μm to about 500 μm from the center of another channel. As shown in Table 3 below, the channel spacing (E) may be optimized to vary the diffusion flux, non-traumatic cell seeding, and volume / footprint of the device.

[0129] Channel 1073 may further be characterized by its aspect ratio, as shown in FIG. 89B. The aspect ratio may be calculated as the ratio between the cell chamber height (A) and the channel diameter (D). The aspect ratio may be at least about 0.5.

Table 3

[0130] As shown in FIG. 107C, the geometric shape of the channel can be designed and optimized in view of its use. Optimal use and cell growth are enabled if the cylindricity, roundness, and perpendicularity of channel 1073 with respect to the first surface of the first membrane 1073 are uniform.

[0131] Figures 108A - B show the inner surface of a typical first membrane of the channel array. As shown in Table 4 below, the surface - to - volume ratio can be increased by the channel spacing and sizing. Figure 90A shows a typical channel array with a channel spacing of 50 μm, and Figure 90B shows a typical channel array with a channel spacing of 270 μm.

[0132] In some embodiments, at least one of the length and width of the channel array device 1070 is from about 0.25 cm to about 3 cm. In some embodiments, each of the plurality of channels 1073 is substantially perpendicular to the first membrane. In some embodiments, the channels 1073 are arranged in a linear array. In some embodiments, the channels 1073 are arranged in a polar array. In some embodiments, the device has a channel number per area along the cross - section greater than about 50 / cm 2 more. In some embodiments, the device has a surface - area - to - volume ratio of at least about 40 cm -1 . In some embodiments, the channel array device 1070 includes a compartment between the first membrane and the second membrane. The compartment may include a single continuous open space. The compartment may have a volume from about 8 μL to about 600 μL.

[0133] In some embodiments, the method of forming a channel array further includes laser - ablating a portion of the first and second membranes within the plurality of channels. In some embodiments, the laser ablation removes the fused portion between the first and second membranes to form an opening.

[0134] In some embodiments, the openings are concentric with the channels to a maximum of 25% of the channel diameter. FIG. 91A (left) shows a detailed image of a typical laser-pierced channel array device. The openings have an unacceptable concentricity with respect to the channels greater than about 25%. FIG. 91A (right) shows a detailed image of a typical laser-pierced channel array device. The openings have an acceptable concentricity with respect to the channels less than about 25%. As shown in FIG. 91C, as the concentricity increases, the uniformity of the width of the fusion region around each opening improves, increasing the bond strength and sealing. Further, as the concentricity increases, the surface area-to-volume ratio increases, reducing the required size of the seal region required for strength and integrity. As the seal surface area decreases, the remaining "shelves" that stimulate the FBR and form a further diffusion distance of the central blood vessel decrease.

[0135] FIG. 92 shows the alignment of the openings and channels from left to right across a typical array device as shown in FIG. 91B. A concentricity value of 0 corresponds to perfect concentricity. For example, the openings formed by the laser ablation parameters associated with Test 894 (center) had a much higher concentricity value than the parameters associated with the ablation parameters associated with Test 898 (right).

[0136] FIG. 93A shows an image of a typical laser-pierced channel array device. FIG. 93B shows an image of the seal interface of the channel array device. FIGS. 94A-B show high and low magnification images of angiogenesis around the implanted channel array device. FIG. 95 shows a diagram of blood vessel-host integration according to some embodiments. In some embodiments, at least one of the first membrane and the second membrane is configured to enable angiogenesis of cells within the device. In some embodiments, at least one of the first membrane and the second membrane is configured to enable angiogenesis of cells within the device in the absence of immunosuppressive therapy.

[0137] The equilibrium state O2 tension distributions with venous capillaries of typical channels at a pressure of 100 mmHg are shown in Fig. 96A for channel diameters of 1,100 μm, 1,300 μm, and 1,500 μm, and for channel edge-to-edge distances of 150 μm and 250 μm. Further, the equilibrium state O2 tension distributions with venous capillaries of typical channels at a pressure of 45 mmHg are shown in Fig. 96A for channel diameters of 1,100 μm, 1,300 μm, and 1,500 μm, and for channel edge-to-edge distances of 150 μm and 250 μm.

[0138] In some embodiments, forming a plurality of channels using a mold includes thermoforming the plurality of channels. Upper perspective and cross-sectional images of a typical non-sintered membrane thermoformed at 360 °C for 7 minutes at a negative pressure of 3 psi are shown in Figs. 115A - B, respectively. As shown, the thermoforming process results in formed channels without cracks or structural collapse. Such a thermoforming process allows for sharp bends and a large cell chamber height of about 558 μm as 1150, resulting in an increased surface area-to-volume ratio and sufficient space for accommodating cells. As shown in Fig. 98, a typical membrane does not rupture when thermoformed, but by using alternative membranes and processes, the fusion strength can be increased by reducing sintering and increasing ductility. Such methods and membranes can further result in obtaining a larger cell chamber height of 1150.

[0139] Fig. 99A shows an array of ruptured (left) and non-ruptured (right) channels. As shown above, when a typical membrane is thermoformed at 3.5 psi and 370 °C for 7 minutes, channels with a maximum cell chamber height are obtained as shown in 118B. As shown in Fig. 100 and Table 6 below, the membrane of 118B enables an excellent cell chamber height of about 672 μm and a cell volume of about 27 ul.

Table 6

[0140] Furthermore, as shown in FIG. 101, when thermoforming at a temperature of about 370° C., the fusion delamination resistance of the membrane can be increased. The channels of this membrane were formed using the fusion tool of FIG. 103A at a temperature of 474° C., a fusion time of 0.05 seconds, and a Z offset of 2.625 mm.

[0141] Furthermore, FIG. 102A shows differential scanning calorimetry graphs for a typical membrane of FIG. 99B starting at a normalized enthalpy of 23.696 J / g with an onset x of 321.25° C. and starting at a normalized enthalpy of 23.141 J / g with an onset x of 321.14° C. Further, FIG. 102B shows electron micrographs of membranes with membrane sintering (right) and without membrane sintering (left). By sintering the membrane, the stability of the cell device can be increased.

[0142] FIGS. 122A and 122B show an explanatory diagram and an image of a typical thermal fusion tool. The thermal fusion tool may include a position-based fusion tool configured to provide a set number of fusion dots (each dot for a set fusion time). In some embodiments, the set fusion time is less than about 1 second. In some embodiments, the fusion tool 1220 dots the first membrane at one or more points for 1 to 6 hours. In some embodiments, the fusion tool 1220 dots the first membrane at one or more points for up to about 16 hours. Further, the temperature of the fusion head can be adjusted to vary the characteristics of the channel and the fusion thereby formed. In some embodiments, the set fusion temperature is from about 250° C. to about 600° C. In some embodiments, the fusion tool 1220 has a dabbing contact area of at least about 0.07 mm 2 is. In some embodiments, the fusion tool 1220 contacts the side of the first membrane facing the thermoformed surface.

[0143] In some embodiments, the channel is formed by placing a first membrane and a second membrane within frame 1221 and dotting one or more points on the first membrane using fusion tool 1220. In some embodiments, the frame surrounds at least a portion of the outer edges of the first membrane and the second membrane. The first membrane and the second membrane may be substantially parallel during fusion. The first membrane and the second membrane may be substantially aligned such that all or most of the second membrane is covered by the first membrane. The first membrane and the second membrane may be separated by a gap distance. In some embodiments, the gap distance is from about 300 μm to about 1,200 μm.

[0144] In some embodiments, when dotting the first membrane, a portion of the first membrane is fused to the second membrane through the first membrane, the second membrane, or both. In some embodiments, at least one of the first membrane and the second membrane is substantially flat. In some embodiments, at least one of the first membrane and the second membrane includes a non-sintered flat sheet.

[0145] Optimizing the fusion dotting parameters may be necessary to prevent misalignments (where fusion is misaligned and inconsistent in some regions) as shown in FIG. 104. The tensile peel strength test results shown in FIG. 105B (measured by a typical machine shown in FIG. 105B) imply that the fusion strength is inversely proportional to the amount of non-parallelism.

[0146] The concentricity of the openings can be improved by the laser drilled openings in the post-fusion device shown in FIGS. 125A - B. However, if there is misalignment in such a laser drilling tool, further discrepancies may occur. As shown in FIG. 97B, such errors can be fixed by an adhesive, but by incorporating a load cell into the fusion probe and using robotic manufacturing and guidance equipment, the manufacturing method can be improved to reduce rework time, waste, and leaks. Incorporating a load cell allows for accurate calibration of the fusion force regardless of the tool shape. When optimized robotic effective fusion parameters are developed, the processing speed is increased and the thermal shrinkage impact is reduced. Using a vision guidance system improves the concentricity of the fusion with respect to the thermoformed channels.

[0147] Alternative typical load-sensitive thermal fusion tools may include a tip, a load cell, and a frame configured to hold a membrane. The thermal fusion tool may include a position-based fusion tool configurable to apply a set fusion force over a set fusion time for a set number of fusion dots. The load cell allows the fusion tool to fuse each point with the same force. A typical array device including a single row of channels formed by the thermal fusion tool is shown in FIG. 107. As shown in FIG. 108, with a minimum fusion time of about 0.05 seconds, the fusion with the strongest peel force was formed between the membranes (.45 N). Since the peel force indicates the fusion strength, a shorter fusion time can increase the stability and lifespan of the device. A complete device formed by the thermal fusion tool at a fusion temperature of 800°F, 1 dot per location, and a fusion force of about 6 pounds is shown in FIG. 109. The round and uniform shape of the channels, as well as the peel force stress-strain curve of the device shown in FIG. 110, confirm a high fusion strength of about 0.45 N.

[0148] A typical channel array device with channels formed by 2, 4, 6, and 8 fusion dots is shown in FIG. 111. As shown in FIG. 112, the strongest fusion was formed by 8 fusion dots, but the strongest fusion with the channels remaining intact was formed by 4 fusion dots.

[0149] Figure 113 shows a bar graph representing the fusion strength of a typical channel array device with a first deformed membrane and a second flat membrane, with the fusion temperature at 800°F, the fusion time at 0.05 seconds, and one fusion dot. Devices with peel forces of approximately 0.4N and approximately 0.36N, which are higher, were obtained at fusion dot forces of 6 and 12 pounds, respectively, but no observable relationship between the two variables was confirmed. Increasing the fusion dot force may not be so ideal because it causes deterioration associated with the fusion tool tip and the membrane frame. This correlation has been confirmed as shown in Figures 138A - B. These figures show that a typical array channel device containing a membrane formed with a fusion force of 8 pounds exhibited membrane cracking, wrinkles, and rectangular or double channels. Therefore, it is a further consideration that the fusion force for a typical membrane is less than 8 pounds.

[0150] Figure 115 shows that when the channels of the device are formed with 4 fusion dots and a fusion force of 6 pounds, the device is stronger (0.65N) than when the channels of the device are formed with 4 fusion dots and a fusion force of 3 pounds (0.47N). Both devices are formed at a fusion temperature of 800°F and a fusion time of 0.05 seconds. Further, Figure 116 shows devices formed at a fusion temperature of 800°F and a fusion time of 0.05 seconds. Devices formed with 4 dots at 6 pounds weight (~0.65) and 2 dots at 8 pounds weight (~0.7N) were stronger than typical devices formed with 1 or 2 dots at 6 pounds weight (~0.47N).

[0151] Figure 117A shows an image of a typical array device with a 3x3 channel array formed simultaneously. Each channel of the plurality of channels is dotted once before one channel is dotted a second time. As shown in Figure 117A, a typical device with a greater peel strength (0.9N) is formed by dotting simultaneously, rather than by fusing each channel by dotting it twice consecutively (0.75N) before dotting another channel.

[0152] A comparison of the peel strength of typical fused membranes is shown in Figure 118 and Table 7 below. Membrane A was formed by a load cell fusion tool that punches 2 times with a force of 8 pounds, Membrane B was formed by a load cell fusion tool that punches 4 times with a force of 6 pounds, and Membrane C was formed by a non-load effective cell fusion tool.

Table 7

[0153] Membranes A and C are shown in Figure 119. The membranes show uniform and consistent fusion. In the case of Membrane A, no significant difference was found in the measured fusion strength over the range of applied fusion forces. In the case of Membrane B, when the fusion force was less than 8 pounds, the strongest membranes with tearing, wrinkles, and low resistance were obtained. Also in the case of Membrane C, the highest fusion strength was measured at 6 pounds. In Figure 121, Membranes A and C are further compared. Membrane C clearly shows a more uniform and higher peel strength.

[0154] Figure 120 shows low-resolution and high-resolution microscopic images of a typical membrane portion fused with 4 fusion punches at a temperature of 800°F, a fusion time of 0.05 seconds, and a fusion force of 6 pounds. These images show that the typical membrane has a fusion strength of approximately 1.38 N, which is equal to the total strength of the non-fused membrane.

[0155] Figure 121 shows a detailed image of the fused channel of a membrane with a seal size of approximately 170+ / -8um. Figure 122A shows a high degree of concentricity between the laser ablation region and the fusion region. Finally, Figure 148 shows the fusion force / peel strength (N) of a typical pre-laser channel array device formed using non-load cell effective (Generation 1) and load cell effective (Generation 2) fusion tools. A typical membrane formed using a load cell effective fusion tool from the membrane shows a significantly higher (1.52 N) peel strength.

[0156] Unless otherwise defined, all technical terms used in this specification have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs.

[0157] As used in this specification, the singular forms "a", "an", and "the" include references to the plural unless the context clearly indicates otherwise. Whenever the term "or" is referred to in this specification, it is intended to include "and / or" unless otherwise specified.

[0158] As used in this specification, the term "about" refers to an amount that is within 10%, 5%, or 1% (including increments therein) of the stated amount.

[0159] As used in this specification, the term "substantially perpendicular" refers to the 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 therein from perpendicular.

[0160] As used in this specification, the term "substantially parallel" refers to the 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 therein from parallel.

[0161] As used in this specification, the phrases "at least one", "one or more", and "and / or" are open-ended expressions that are both conjunctive and disjunctive in use. For example, 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" each mean A alone, B alone, C alone, A and B together, A and C together, B and C together, or A, B, and C together.

[0162] Preferred embodiments of the present disclosure have been illustrated and described herein. As will be apparent to those skilled in the art, such embodiments are shown by way of example only. Many variations, modifications, and alternatives will occur to those skilled in the art without departing from the present disclosure. Of course, various alternatives to the embodiments of the present disclosure described herein may be used when practicing the present disclosure. It is intended that the scope of the present disclosure be defined in the following claims and that the methods and structures fall within the scope of these claims and their equivalents.

Example

[0163] The present disclosure is further limited by the following non-limiting examples.

[0164] Example 1 Hexagonal Channel Array Device for Evaluation in Small Animals In this example, the configuration of a hexagonal channel array device for evaluation in small animals will be described. FIG. 6 shows renderings of hexagonal channel array devices of various sizes. The channel array device designed for testing in rats is an elongated hexagonal shape with dimensions of 1.9 cm x 0.8 cm. The rat device has 93 channels, the SA:V ratio is 82 cm -1 and the safety volume is 97%, and it can hold a volume of 43 μl. The channel array device designed for testing in mice is hexagonal in shape with dimensions of 0.64 cm x 0.55 cm. The mouse device has 19 channels, the SA:V ratio is 82 cm -1 and the safety volume is 95%, and it can hold a volume of 10 μl. The mid device is 1.4 cm x 0.55 cm, has 49 channels, and the SA:V ratio is 83 cm -1 and the safety volume is 96%, and it can hold a volume of 24 μl. The size of the device can be scaled while maintaining a constant SA:V ratio.

[0165] Example 2 Ultra-Fine Structure Analysis of PVDF Membrane after Deformation Step In this example, the ultra - fine structure analysis of the PVDF membrane after the deformation step will be described. The PVDF membrane underwent a deformation step under a combination of pressure and temperature. The combinations were 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, and 140 psi and 165 °C. Figures 12 and 16 show scanning electron micrographs of cross - sectional views of the PVDF membrane after the deformation step. Figure 17 shows the measured channel depth or feature depth (μm) for the cell - containing device when the deformation step was performed under different pressure and temperature conditions. The PVDF membrane has different channel depths depending on the temperature and pressure conditions, and generally the channels are deeper at a high temperature of 165 °C. The dimensions of the channels can be controlled by the temperature and pressure conditions. The control of the channel dimensions is important for determining the SA:V ratio of the cell - containing device.

[0166] Example 3 Analysis of the PVDF cell - containing device after the fusion step In this example, the analysis of the PVDF cell - containing device after the fusion step will be described. The first PVDF membrane underwent a deformation step under 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. Next, the deformed membrane was fused to a second PVDF membrane at 225 °C. Figure 13 shows a scanning electron micrograph of a cross - sectional view of the PVDF cell - containing device after the fusion step. The first membrane was deformed at 65 psi and 165 °C, 100 psi and 150 °C, 140 psi and 135 °C, 140 psi and 150 °C and fused to the second membrane. The first membrane deformed outside these temperature and pressure parameters showed insufficient fusion, and the seam between the first and second membranes was clearly visible in the scanning electron micrograph.

[0167] Figure 19 shows a scanning electron microscope photograph of a cross-section of a cell containment device with insufficient fusion. The deformation step is performed at various pressures and temperatures. The heat flow measurement of the membrane deformed at 165 °C and 140 psi showed a secondary peak at approximately 175 °C when measured by DSC. This indicates the rearrangement of the crystals in this deformed membrane. The heat flow measurement of the membrane deformed at 150 °C and 100 psi, which successfully fused to the second membrane, did not show a secondary peak.

[0168] Example 4 DSC Analysis of Fusion and Non-Fusion Conditions In this example, the DSC analysis of fusion and non-fusion conditions will be described. The first PVDF membrane was subjected to a deformation step under 100 psi and 160 °C or 100 psi and 173 °C and subsequently quenched. Figure 20 shows that the heat flow measurement of the membrane deformed at 100 psi and 160 °C had a small shoulder peak and was similar to the baseline PVDF membrane. That is, this membrane preserved its crystal arrangement and showed that it fused with the second membrane. The heat flow measurement of the membrane deformed at 100 psi and 173 °C had a secondary peak. That is, this membrane formed a crystal region and showed that it did not fuse with the second membrane.

[0169] Example 5 Surface Profile of the Cell Containment Device In this example, the surface profile of the cell containment device will be described. Figure 18 shows the surface interferometry of a cell containment device that had a smooth surface with a uniform geometric shape within its channel array. The active region had a smooth porous surface, while the fusion region had a flat surface.

[0170] Example 6 Ultrastructural Analysis of ePTFE after Deformation and Fusion Steps In this example, the ultrastructural analysis of the ePTFE membrane after the deformation and fusion steps will be described. The ePTFE membrane was subjected to the deformation step at 30 psi and 340 °C, 4 psi and 360 °C, or 6 psi and 360 °C. Figure 19 shows a scanning electron micrograph of the ePTFE membrane after the deformation step. The ePTFE membrane was able to form channels. Generally, as the temperature increased, an increase in deformation and a decrease in nodes were observed. Figure 20 shows the fused ePTFE membrane that was subjected to the deformation step at 6 psi and 360 °C and the fusion step at 370 °C for 5 minutes. The ePTFE membrane requires different temperature and pressure ranges for the deformation and fusion steps compared to the PVDF membrane. The manufacturing steps described here may be applied to membranes of different materials and succeed in forming a cell containment device.

[0171] Example 7 In Vivo Implantation of the Cell Containment Device into Rats In this example, the in vivo transplantation of cell containment devices into rats will be described. These devices had the same channel diameter of approximately 350 μm, but different channel spacings. The low-density device had a channel spacing of approximately 500 μm between channels, and the high-density device had a channel spacing of approximately 200 μm. As shown in FIG. 28, cell containment devices with high and low channel densities were transplanted into the in vivo of various locations of normal rats (preperitoneal, omental, suprahepatic, and subcutaneous transplantation). FIG. 29 shows angiogenesis around the cell containment device after preperitoneal transplantation. Microvessels were observed in each channel within the device. The H&E histological image of the vasculature showed the presence of smooth muscle cells, as seen in arteries. FIG. 37 shows further angiogenesis and further branching observed in the high-channel density device compared to the low-channel density device. FIG. 32 shows the count of blood vessels and branches per device for the low-channel density and high-channel density devices. The high-channel density device had approximately 1000 blood vessels per device compared to approximately 500 blood vessels per device in the case of the low-channel density device. Also, the high-channel density device had significantly more branches, approximately 500 branches per device, compared to approximately 300 branches per device in the case of the low-channel density device. The higher the channel density of the device, the more angiogenesis around the device. The advantage of increased angiogenesis seen in the higher-channel devices is that, since more channels are added to the device, it may be necessary to balance the integrity of the device. Channel density may affect the level of angiogenesis around the cell containment device.

[0172] For in vivo transplantation, the cell containment devices were filled with cells. FIG. 42 shows H&E-stained tissue sections in which cells completely filled the continuous internal spaces of various cell containment devices. In the device (upper) formed using a 20-μm thick ePTFE membrane 5310, cells 5320 filled the entire single continuous internal space. This device had a modeled insulin diffusion rate of approximately 11 ng / cm per 10 minutes 2There may be cases. As shown below, a device formed using a 125-μm thick PVDF membrane 5330 was filled with cells 5320 throughout its single continuous internal space. The modeled insulin diffusion rate of this device was about 6 ng / cm per 10 minutes 2 There may be cases. Figure 43 shows an image of an ePTFE cell-containing device filled with cells up to its maximum capacity. In the upper panel, a single continuous internal space of the device filled with cells throughout the device is shown by an H&E-stained tissue section. In the lower left, a close-up image shows a microstructure containing cells within one pocket of the internal space with a height of about 150 μm and a width of about 400 μm. In the lower right, an optical micrograph at 5x magnification of the ePTFE cell-containing device is shown. The continuous filling of the internal space of the fabricated cell-containing device with channels is shown. This demonstrates that the entire internal space of the cell-containing device can be filled with cells.

[0173] The cell containment device was transplanted into the living body of a rat for a long time, and the cells in the cell containment device survived during the long-term in-vivo transplantation. Figures 26 and 27 show the vascular structures around the cell containment device and passing through the channels 20 days (Figure 26) and 90 days (Figure 27) after transplantation into the preperitoneal site in nude rats. Figure 44 shows an H&E-stained tissue section of the PVDF cell containment device 5510 with cells 5520 90 days after in-vivo transplantation into the preperitoneal site of a rat. The SC pancreatic islet cells 5520 in the device 5510 transplanted into the preperitoneal site had high cell contents on day 90, and angiogenesis 5530 was visible around the device and through the channels. The host tissue formed new tissue around the device and through the channels of the device. Figure 45 shows an H&E-stained tissue section of the PVDF cell containment device 5610 filled with SC pancreatic islet cells 5620 90 days after transplantation into the subcutaneous and preperitoneal sites of a nude rat capable of a foreign body reaction. The SC pancreatic islet cells 5620 in the device 5610 transplanted into both the subcutaneous and preperitoneal sites had high cell contents on day 90. This indicates that the SC pancreatic islet cells survived in the device for 90 days at both transplantation sites. Angiogenesis 5630 was observed around the device and through the channels of the device transplanted into both the subcutaneous and preperitoneal sites. Some of the angiogenesis appeared to have arteriole-like features and defined a vascular wall with a thickness of multiple cell layers. The outer surface of the device appeared to integrate with the host tissue at both transplantation sites. The density of fibrosis was higher around the device at the subcutaneous site, but there was no obvious effect on the survival of the cells in the device (upper and lower left). The host tissue also existed through the channels of the device, providing support for the vascular bundle tissue and further mechanical stability for the device. This indicates that cells can be transplanted into the cell containment device at multiple transplantation sites, maintaining high cell viability of the cells in the device over a long time, and angiogenesis and new tissue can be formed around and through the device.

[0174] Example 8 Effect on the design of the cluster size In this example, it will be explained how the cluster size can affect the design. The cluster size refers to the size of the cell aggregate filled in the device.

[0175] Example 9 Deformation conditions of ePTFE membrane In this example, the results of the deformation conditions for the ePTFE membrane are shown. Figure 41 shows a cross-sectional image of an ePTFE cell-containing device formed by thermal deformation at 360 °C and 6 psi (upper part during formation), and then hard-cast using resin (lower part). The ePTFE membrane during formation did not seem to retain its channel shape and did not seem to have more small fiber structures than nodes. The ePTFE membrane hard-cast using resin seemed to retain its channel shape after the deformation step.

[0176] Example 10 T-peel test of two flat ePTFE membranes In this example, the T-peel tests of sintered and non-sintered ePTFE membranes that test the bond strength of the fused ePTFE membrane will be described. Figure 43 shows the breaking load (N) by the ASTM T-peel test according to the ASTM D882-08 standard for the thin film tensile test of two flat ePTFE membranes fused for different lengths of time (1, 5, 15 seconds) at different temperatures. The ePTFE membranes were either sintered at 370 °C for 7 minutes (AS) or non-sintered (AU) before fusion. The sintered ePTFE membrane had a fusion temperature of about 320 °C to 325 °C, and the non-sintered ePTFE membrane had a fusion temperature of about 340 °C to 350 °C (measured by DSC). Two flat ePTFE membranes were fused to each other for 1, 5, or 15 seconds at various temperatures in the range of 302 °C to 427 °C. After undergoing the ASTM T-peel test, the breaking or failure load (N) was recorded. Generally, the fusion between sintered-sintered membranes (AS / AS) had the lowest breaking load (in the range of near 0 N to about 0.3 N), compared to non-sintered-non-sintered membranes (AU / AU) (in the range of near 0 N to about 1 N), or non-sintered-sintered membranes (AU / AS) (in the range of about 0.2 N to 0.7 N). Generally, the fusion of AU / AU membranes had a higher breaking load. Generally, the breaking load increased with the increase in the fusion temperature. Generally, having at least one non-sintered membrane resulted in a higher breaking load.

[0177] Example 11 T-peel test of ePTFE device In this example, the T-peel test of the ePTFE device will be described. Figure 38 shows the tool used for the ASTM T-peel test according to the ASTM D882-08 standard for thin film tensile tests. Using this tool, the fused cell-containing ePTFE device was tested. The tested device at the time of fracture is shown. The graph shows the stress-strain curve of the ePTFE device. The first membrane was sintered and deformed, and the second membrane was flat and unsintered, and these were fused at 474 °C for 0.05 seconds. The ePTFE device had a fracture strain greater than 60% and reached a load of approximately 0.4 N. This was approximately 78% of the load of the fused flat membrane with a wider fusion area. This stress-strain curve indicated that the fusion site was fused and remained fused even at high tensile strains.

[0178] Example 12 Burst pressure of the ePTFE device In this example, the measurement of the burst pressure of the cell-containing device will be described. To test the seal strength of the device, the device was filled with water at 1 psi per 10 seconds, and the pressure at fracture or burst pressure was measured. Figure 39 shows a graph of the filling pressure (psi) at fracture for an ePTFE cell-containing device without a filled frame for the burst pressure test and an ePTFE cell-containing device (eCAD) prototype. In the case of the PVDF prototype, the burst pressure at fracture was approximately 10 psi. In the case of the ePTFE prototype, the burst pressure at fracture was approximately 11 psi. This burst pressure is much higher than the maximum filling pressure of approximately 2 psi to which the device can be exposed under typical filling conditions.

[0179] Example 13 Cell-containing device prototype In this example, the prototype of the cell-containing device will be described. Figure 14 shows a rendering of the cell-containing device with a scalloped perimeter and variations in channel dimensions to achieve various SA:V ratios. As shown in Figure 14, the cell-containing device is characterized by an overall height 1801, an internal height 1802, a membrane thickness 1803, an internal spacing 1804, an inner diameter 1805, a through-hole inner diameter 1806, and a through-hole spacing 1807.

[0180] In some embodiments, the overall height 1801 is from about 400 μm to about 1,600 μm. In some embodiments, the overall height 1801 is at least about 400 μm. In some embodiments, the overall height 1801 is at most about 1,600 μm. In some embodiments, the overall height 1801 is from about 400 μm to about 600 μm, from about 400 μm to about 850 μm, from about 400 μm to about 1,000 μm, from about 400 μm to about 1,200 μm, from about 400 μm to about 1,400 μm, from about 400 μm to about 1,600 μm, from about 600 μm to about 850 μm, from about 600 μm to about 1,000 μm, from about 600 μm to about 1,200 μm, from about 600 μm to about 1,400 μm, from about 600 μm to about 1,600 μm, from about 850 μm to about 1,000 μm, from about 850 μm to about 1,200 μm, from about 850 μm to about 1,400 μm, from about 850 μm to about 1,600 μm, from about 1,000 μm to about 1,200 μm, from about 1,000 μm to about 1,400 μm, from about 1,000 μm to about 1,600 μm, from about 1,200 μm to about 1,400 μm, from about 1,200 μm to about 1,600 μm, or from about 1,400 μm to about 1,600 μm. In some embodiments, the overall 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, the internal height 1802 is from about 300 μm to about 1,200 μm. In some embodiments, the internal height 1802 is at least about 300 μm. In some embodiments, the internal height 1802 is at most about 1,200 μm. In some embodiments, the internal height 1802 is from about 300 μm to about 400 μm, from about 300 μm to about 650 μm, from about 300 μm to about 800 μm, from about 300 μm to about 1,000 μm, from about 300 μm to about 1,200 μm, from about 400 μm to about 650 μm, from about 400 μm to about 800 μm, from about 400 μm to about 1,000 μm, from about 400 μm to about 1,200 μm, from about 650 μm to about 800 μm, from about 650 μm to about 1,000 μm, from about 650 μm to about 1,200 μm, from about 800 μm to about 1,000 μm, from about 800 μm to about 1,200 μm, or from about 1,000 μm to about 1,200 μm. In some embodiments, the internal height 1802 is about 300 μm, about 400 μm, about 650 μm, about 800 μm, about 1,000 μm, or about 1,200 μm.

[0181] In some embodiments, the membrane thickness 1803 is from about 50 μm to about 250 μm. In some embodiments, the membrane thickness 1803 is at least about 50 μm. In some embodiments, the membrane thickness 1803 is at most about 250 μm. In some embodiments, the membrane thickness 1803 is from about 50 μm to about 75 μm, from about 50 μm to about 100 μm, from about 50 μm to about 125 μm, from about 50 μm to about 150 μm, from about 50 μm to about 175 μm, from about 50 μm to about 200 μm, from about 75 μm to about 100 μm, from about 75 μm to about 125 μm, from about 75 μm to about 150 μm, from about 75 μm to about 175 μm, from about 75 μm to about 200 μm, from about 100 μm to about 125 μm, from about 100 μm to about 150 μm, from about 100 μm to about 175 μm, from about 100 μm to about 200 μm, from about 125 μm to about 150 μm, from about 125 μm to about 175 μm, from about 125 μm to about 200 μm, from about 150 μm to about 175 μm, from about 150 μm to about 200 μm, or from about 175 μm to about 200 μm. In some embodiments, the membrane 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.

[0182] In some embodiments, the internal spacing 1804 is from about 40 μm to about 500 μm. In some embodiments, the internal spacing 1804 is at least about 40 μm. In some embodiments, the internal spacing 1804 is at most about 500 μm. In some embodiments, the internal spacing 1804 is from about 40 μm to about 60 μm, from about 40 μm to about 80 μm, from about 40 μm to about 100 μm, from about 40 μm to about 150 μm, from about 40 μm to about 200 μm, from about 40 μm to about 270 μm, from about 40 μm to about 350 μm, from about 40 μm to about 400 μm, from about 40 μm to about 500 μm, from about 60 μm to about 80 μm, from about 60 μm to about 100 μm, from about 60 μm to about 150 μm, from about 60 μm to about 200 μm, from about 60 μm to about 270 μm, from about 60 μm to about 350 μm, from about 60 μm to about 400 μm, from about 60 μm to about 500 μm, from about 80 μm to about 100 μm, from about 80 μm to about 150 μm, from about 80 μm to about 200 μm, from about 80 μm to about 270 μm, from about 80 μm to about 350 μm, from about 80 μm to about 400 μm, from about 80 μm to about 500 μm, from about 100 μm to about 150 μm, from about 100 μm to about 200 μm, from about 100 μm to about 270 μm, from about 100 μm to about 350 μm, from about 100 μm to about 400 μm, from about 100 μm to about 500 μm, from about 150 μm to about 200 μm, from about 150 μm to about 270 μm, from about 150 μm to about 350 μm, from about 150 μm to about 400 μm, from about 150 μm to about 500 μm, from about 200 μm to about 270 μm, from about 200 μm to about 350 μm, from about 200 μm to about 400 μm, from about 200 μm to about 500 μm, from about 270 μm to about 350 μm, from about 270 μm to about 400 μm, from about 270 μm to about 500 μm, from about 350 μm to about 400 μm, from about 350 μm to about 500 μm, or from about 400 μm to about 500 μm. In some embodiments, the internal 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.

[0183] In some embodiments, the inner diameter 1805 is from about 300 μm to about 1,600 μm. In some embodiments, the inner diameter 1805 is at least about 300 μm. In some embodiments, the inner diameter 1805 is at most about 1,600 μm. In some embodiments, the inner diameter 1805 is from about 300 μm to about 500 μm, from about 300 μm to about 700 μm, from about 300 μm to about 900 μm, from about 300 μm to about 1,100 μm, from about 300 μm to about 1,300 μm, from about 300 μm to about 1,600 μm, from about 500 μm to about 700 μm, from about 500 μm to about 900 μm, from about 500 μm to about 1,100 μm, from about 500 μm to about 1,300 μm, from about 500 μm to about 1,600 μm, from about 700 μm to about 900 μm, from about 700 μm to about 1,100 μm, from about 700 μm to about 1,300 μm, from about 700 μm to about 1,600 μm, from about 900 μm to about 1,100 μm, from about 900 μm to about 1,300 μm, from about 900 μm to about 1,600 μm, from about 1,100 μm to about 1,300 μm, from about 1,100 μm to about 1,600 μm, or from about 1,300 μm to about 1,600 μm. In some embodiments, the 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.

[0184] In some embodiments, the through-hole inner diameter 1806 is from 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 from about 100 μm to about 200 μm, from about 100 μm to about 300 μm, from about 100 μm to about 400 μm, from about 100 μm to about 500 μm, from about 100 μm to about 600 μm, from about 200 μm to about 300 μm, from about 200 μm to about 400 μm, from about 200 μm to about 500 μm, from about 200 μm to about 600 μm, from about 300 μm to about 400 μm, from about 300 μm to about 500 μm, from about 300 μm to about 600 μm, from about 400 μm to about 500 μm, from about 400 μm to about 600 μm, or from 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.

[0185] In some embodiments, the through-hole pitch 1807 is from about 100 μm to about 400 μm. In some embodiments, the through-hole pitch 1807 is at least about 100 μm. In some embodiments, the through-hole pitch 1807 is at most about 400 μm. In some embodiments, the through-hole pitch 1807 is from about 100 μm to about 200 μm, from about 100 μm to about 300 μm, from about 100 μm to about 400 μm, from about 200 μm to about 300 μm, from about 200 μm to about 400 μm, or from about 300 μm to about 400 μm. In some embodiments, the through-hole pitch 1807 is about 100 μm, about 200 μm, about 300 μm, or about 400 μm.

[0186] In one example, the channel 1810 of the device may have an inner diameter 1805 of 800 μm, an internal height 1802 of 650 μm, and a through-hole inner diameter 1806 of the through-hole 1820 of 300 μm. This device may have an overall height 1801 of 850 μm, a 100-μm membrane thickness 1803, and an internal spacing 1804 between channels 1810 of 270 μm. This device has a SA:V ratio of 77 cm -1It may also be. In another example, the channel 1810 of the device may have an inner diameter 1805 of 650 μm, an internal height 1802 of 650 μm, and a through-hole inner diameter of the through-hole 1820 of 400 μm. This device may have an overall height 1801 of 850 μm, a 100-μm membrane thickness 1803, and an internal spacing 1804 between the channels 1810 of 80 μm. This device may have an SA:V ratio of 138 cm -1 It may also be. To achieve different SA:V ratios, the dimensions and spacing of the channels 1810 may be adjusted. The device may have a through-hole spacing 1807 of about 200 μm.

[0187] Example 14 Human device prototype In this example, a prototype of a cell-containing device assembled on a frame for human use will be described. FIG. 40 shows a prototype of three cell-containing devices assembled on a frame for human use. The cell-containing device may have a hexagonal shape. The frame may have dimensions of about 26.9 mm x 75.6 mm and may have an oval shape with perforations in the space between the cell-containing devices. The cell-containing device may be about 850 μm thick and have a footprint of 20 cm 2 It may also be. The assembled device may hold about 400 million cells and have a cell mass volume of about 350 μL. These cells may be able to form insulin.

[0188] Example 15 Surface modification of the membrane In this example, a process for modifying the membrane surface will be described. The membrane may be treated to impart hydrophilicity onto a membrane with a hydrophobic surface. The surface of the membrane may be modified by crosslinking a hydrophilic polymer. The polymer may also be biocompatible, and a biocompatible coating may be formed on the membrane. In one example, in a thermal-initiated polymerization process using an ammonium persulfate (APS) initiator, hydroxypropyl acrylate (HPA) may be crosslinked on the membrane using tetra(ethylene glycol) diacrylate (TEGDA). FIG. 41 shows an example of a protocol for forming a hydrophilic coating on the surface of the membrane. The ePTFE device was immersed in 100% ethanol and then in 30% ethanol for 3 minutes. The device was immersed in 3% HPA, 2% TEGDA, 1% APS in 30% ethanol for 5 minutes and heated from room temperature to 80 °C. Then, the device was boiled in 100% ethanol for 5 minutes, immersed in Milli-Q water or ultrapure water for 30 minutes, and dried. FIG. 38 shows the ePTFE cell containment device after immersion in a hydrophilic coating treatment in water. FIG. 41 shows an electron micrograph of the ePTFE device after surface modification.

[0189] Example 16 Dot diameter and density of the device In this example, the dot diameter and density (e.g., the spacing between centers) of the device are described. Adhesive dots were stacked in two layers. The device was manually flipped to form a pattern of adhesive dots that act to limit the thickness of the device during cell filling. A pattern of 37 dots (each dot having a diameter of approximately 1.25 mm and a center-to-center spacing of 3.3 mm) was deposited in two consecutive layers. First, the pattern was created on the membrane by applying the adhesive for 0.05 seconds and then curing each dot individually for 1 second. A smaller second layer was applied over the first pattern with a dispense time of 0.03 seconds and left uncured. Next, the peripheral adhesive was placed as described herein, and the membrane was placed over the uncured adhesive using machine vision. After a penetration time of 1 second, curing of the entire assembly was performed by first curing the periphery and then sweeping the interior region to cure the dot pattern for 28 and 112 seconds, respectively. Next, the completed device was manually removed from the assembly platform and placed in a secondary container, and post-assembly heat curing was performed at 37°C for 2 hours.

[0190] Example 17 Dot diameter and filling parameters of the device In this example, the filling volume of devices with dots of various configurations will be described. The dot diameter and density (e.g., the spacing between centers) can affect the acceptable filling volume by forming a series of columns. The devices were prepared as described in Example 16 using dots of various configurations. The devices were prepared using a dot pattern. Each dot had a diameter of approximately 1.25 mm, and the dot pitch was 2.6 mm, 3.3 mm, or 4.4 mm. The center of one dot was placed at a dot pitch distance from the center of its adjacent dot. For devices of the same dimensions, a device with a shorter dot pitch may have a higher dot density than a device with a longer dot pitch. For devices with the same dot diameter and device dimensions, a device with a shorter dot pitch may have a smaller internal volume available for filling than a device with a longer dot pitch. Then, the device was filled with cell suspension either without being restricted on the outside of the device or with an external restricting part. The external restricting part may prevent the membrane of the device from expanding outward so that the membranes are separated from each other. Figure 2 shows how the amount of cells filled in the device changes with the dot pitch and the presence of the restricting part. When the dot pitch decreased, the amount of cells to be filled was small. The amount of cells decreased from 108x10 6 cells in a device filled without using a restricting part with a dot pitch of 4.4 mm to 42x10 6 cells in a device filled using an external restricting part with a dot pitch of 2.6 mm. For devices with the same dot pitch, a large amount of cells filled in the device decreased when filled using an external restricting part.

[0191] Example 18 Adhesive dots on the cell-containing device When forming dots on the cell containment device, the device can be manually flipped to form a pattern of adhesive dots that act to limit the thickness of the device during cell filling. For example, a pattern of 37 dots (each dot having a diameter of approximately 1.25 mm and a center-to-center spacing of 3.3 mm) was deposited in two consecutive layers. First, the adhesive was applied by a 0.05 second coating and each dot was cured separately for 1 second to create the pattern on the membrane. A smaller second layer was applied over the first pattern with a dispense time of 0.03 seconds and left uncured. Next, peripheral adhesive was placed on the membrane and the membrane was placed over the uncured adhesive using machine vision. After a 1 second dwell time, curing of the entire assembly was performed by first curing the perimeter and then sweeping the interior region to cure the dot pattern for 28 and 112 seconds, respectively. Next, the completed device was manually removed from the assembly platform and placed in a secondary container, and post-assembly heat curing was performed at 37 °C for 2 hours.

[0192] Example 19 In Vivo Implantation of Ultra-Thin Devices In this example, transplantation of an ultrathin cell-containing device into a NODscid gamma (NSG) mouse model, an immunodeficient mouse model, will be described. Diabetes was induced in the NSG mice. As shown in Figure 61, the blood glucose level of the mice rose above 400 mg / dL. After inducing diabetes, the ultrathin device was transplanted into the epididymal fat pad of the mice. Figure 61A shows an example of the transplanted ultrathin device. These transplanted ultrathin devices contain a hydrophilic coating and a high-flux ePTFE membrane with 8 million SC pancreatic islet cells. After transplanting the ultrathin device, the blood glucose levels of all test mice decreased to about 100 mg / dL, approaching the levels before diabetes induction, and remained so until the ultrathin device was removed or explanted for 90 days, as shown in Figure 61. The explanted ultrathin device was analyzed histologically. Figures 62B and C show a high density of cells throughout the device and the nuclei of the device after transplantation into the NSG mouse model for 90 days. An ultrathin device filled with pancreatic islet cells can be transplanted into a diabetic subject to reduce and normalize the subject's blood glucose level over a long period of time.

[0193] Example 20 In Vivo Transplantation of the AS1 Ultrathin Device In this example, the state of the cells in the ultrathin device after in vivo transplantation into a mouse model for 30 days and 3 months will be described. Figure 62 shows a low-magnification image of a tissue section of an ultrathin device filled with cells after in vivo transplantation into a mouse model for 30 days. The ultrathin device was formed using an AS1 membrane and filled with SC pancreatic islet cells (designated as SEM-01). Figure 63 shows a high-magnification image of the tissue section. The cells were well-distributed and viable throughout the device after 30 days of in vivo.

[0194] Example 21 In Vivo Transplantation of Endocrine Cells in the Ultrathin Device In this example, the cell phenotype of endocrine cells in an ultrathin device after 3 months of in vivo transplantation in a mouse model will be described. Figure 64 shows microaggregates of endocrine cells before encapsulation and filling in the cell containment device. Figure 65 shows a stained tissue image of an ultrathin device filled with microaggregates of endocrine cells after 3 months of in vivo transplantation in a mouse. The blue stain indicates the nucleus, the orange-brown stain indicates the presence of C-peptide, and the pink stain indicates the presence of glucagon. The orange-brown and pink stains in the microaggregates of Figure 64 indicate the presence of active endocrine cells that secrete C-peptide and glucagon. The orange-brown and pink stains in the microaggregates of Figure 65 indicate that the endocrine cells in the ultrathin device remained viable and active during the 3 months of in vivo transplantation and maintained their endocrine gland phenotype, secreting C-peptide and glucagon.

[0195] Example 22 Intraperitoneal glucose tolerance test using an ultrathin device In this example, an intraperitoneal glucose tolerance test using ultrathin devices of various configurations will be described. Figure 66 shows the levels of serum C-peptide and total insulin content in mice transplanted with endocrine cells or ultrathin devices filled with endocrine cells. The test groups were D601mcRA2 endocrine cells transplanted under the capsule (Group A), ultrathin devices filled with D601 cells with an AS1 membrane (Group B), and ultrathin devices filled with D601 cells with a type A ePTFE membrane (Group C). Cells or devices were transplanted into mice and an intraperitoneal glucose tolerance test was performed. Serum C-peptide levels were measured at baseline (blue) and 30-minute glucose stimulation (orange), along with the total insulin content in the graft. All groups showed an increase in serum C-peptide levels with glucose stimulation, indicating insulin production with glucose stimulation. Group A had a serum C-peptide level of approximately 100 pM at baseline and approximately 250 pM with glucose stimulation, and a total insulin content of approximately 700 μg. Group B had a serum C-peptide level of approximately 100 pM at baseline and approximately 200 pM with glucose stimulation, and a total insulin content of approximately 500 μg. Group C had a serum C-peptide level of approximately 50 pM at baseline and approximately 600 pM with glucose stimulation, and a total insulin content of approximately 550 μg. In this example, as measured by the increase in serum C-peptide levels, evidence of glucose control by increased insulin production is shown.

[0196] Example 23 Host / Ultrathin Device Interaction in Nude Mouse Model In this example, the interaction between the host and the ultrathin device after device transplantation into a nude mouse model will be described. Figure 67 shows an H&E stained image of an explant of an ultrathin device containing a coated selective permeable membrane filled with SC pancreatic islet cells at the preperitoneal site of a nude rat 3 months later. The image shows the alleviation of the foreign body reaction (FBR) in the ultrathin device with a selective permeable membrane having a hydrophilic coating due to the absence of macrophages and macrophage fusion, and angiogenesis. The image also shows the separation of host tissue from the inside of the ultrathin device and the concentration of viable cells in the device.

[0197] Example 24 Cell viability and phenotype in a transplanted device filled with SC pancreatic islet cells In this example, cell viability and phenotype maintenance after 12-week transplantation of an ultrathin device filled with SC pancreatic islet cells into a nude mouse model are described. Figure 68 shows an H&E stained image of an ultrathin device filled with 16 million SC pancreatic islet cells after 12-week transplantation into the preperitoneal site of a nude mouse model. The image shows a high level of cell viability in the device nucleus after 12 weeks in vivo. Figure 69 shows serum C-peptide levels over 60 minutes in four different mice transplanted with an ultrathin device filled with SC pancreatic islet cells after glucose administration into the peritoneal cavity. Serum C-peptide increased over time from approximately 30 - 100 pM at 0 minutes to over approximately 1000 pM in the four mice. This indicates that the SC pancreatic islet cells in the ultrathin device transplanted into the mice are active and maintain their ability to produce insulin.

[0198] Example 25 Biocompatibility of an ultrathin device transplanted into an immunocompetent mouse model In this example, the biocompatibility of an ultrathin device with an AS1 membrane transplanted into an immunocompetent C57BL / 6 mouse model is described. An empty mouse-sized ultrathin device with a coated AS1 membrane was placed subcutaneously in C57BL / 6 mice to evaluate the baseline host response to the materials used for the ultrathin device. After 1 month, the device was evaluated for maintenance of device integrity from host tissues and cells and for the foreign body reaction (FBR). Figures 70A and 70B show no cells inside the device and no FBR. The FBR is described as peaking within the first month (Beets, 1998). The images show that the device integrity was maintained while in vivo, and that the device materials and coating are biocompatible and do not induce an FBR.

[0199] Example 26 Transplantation of an ultrathin device into a diabetic NSG mouse model In this example, transplantation of an ultrathin device encapsulating rat pancreatic islet cells with an AS1 membrane into a diabetic NSG mouse model will be described. An ultrathin device filled with 400 IEQ rat pancreatic islet cells with an AS1 membrane was transplanted into diabetic NSG mice for 90 days. Figure 71 shows an H&E stained image of the ultrathin device with viable intact rat pancreatic islet cells after 90 days in vivo. Figure 72 shows the blood glucose levels before transplantation of the ultrathin device and over 90 days after transplantation. After inducing diabetes, insulin pellets were administered to the animals to control hyperglycemia for the first 10 - 20 days of the experiment. After transplantation of the ultrathin device (labeled "transplantation"), over 90 days of transplantation, the blood glucose levels decreased from 400 mg / dL or more to about 100 mg / dL to about 300 mg / dL. The blood glucose levels increased after explanting the device from the mice. This indicates that long-term glucose control can be achieved by transplantation of the ultrathin device.

[0200] Example 27 Cell filling of the ultrathin device Figures 87 and 88 show ultrathin devices of various configurations. The ultrathin device designed for transplantation into a mouse model can be filled with 8 million cells and has a fused dot at the center of the device. The ultrathin device designed for human transplantation can neither have a fused dot nor an array of dots and can be filled with 133 million cells.

[0201] Example 28 Mass flow rate of the ultrathin device with dots FIG. 75A shows an ultra-thin device for human transplantation with an array of spot welds that provide an adhesion limiting portion that prevents the two membranes of the device from bending away from each other when the device is filled. FIG. 75B shows a setup using a porous metal platen that provides an external limiting portion for an ultra-thin device that suppresses bending of the two membranes of the device away from each other when the device is filled. An external porous limiting portion may be used when filling any cell-containing device (e.g., an ultra-thin device and an ultra-thin device with spot welds). FIG. 77 shows the mass flow rate measured from the filling of an ultra-thin device for human transplantation with or without an external porous limiting portion with a 3.3 mm dot pitch. The mass flow rate (measured in cm 3 / min or standard cubic centimeters per minute (sccm)) was similar with or without an external porous limiting portion, reaching a peak of about 7.5 or 8 sccm in about 10 seconds and decreasing with time. After reaching the first peak, the mass flow rate was slightly higher without an external porous limiting portion. FIG. 78 shows an H&E stained image of the cell distribution across an ultra-thin device with an adhesion limiting portion, similar to the device shown in FIG. 75A. This shows that the device with an adhesion limiting portion can be uniformly filled across the entire device.

[0202] Example 29 Cell Filling of Ultra-Thin Devices With or Without Dots In this example, the cell filling of ultrathin devices with or without dots will be described. Figures 79A and B show two configurations of hexagonal ultrathin devices: without dots (A) and with a 3.3 mm dot array matrix throughout the device (B). Figure 80 shows the amount of cells that can fill a single ultrathin device when there are no dots as in the case of Figure 79A and when there is a 3.3 mm dot array matrix as in the case of Figure 79B. The device without dots could be filled with approximately 120 million cells, and the device with a 3.3 mm dot matrix could be filled with approximately 80 million cells. This indicates that the cell filling can be adjusted using the dot array matrix and that the membrane of the device can be prevented from expanding or curving during cell filling.

[0203] Example 30 Cell Filling of Ultrathin Devices with a Limiting Portion In this example, the cell filling of ultrathin devices with or without a porous limiting portion will be described. Figures 81A and B show two configurations of hexagonal ultrathin devices with a 3.3 mm dot array matrix: filled without any limiting portion (A) and filled with a porous platen spaced apart using a 400 μm spacer (B). Figure 82 shows the amount of cells that can fill a single ultrathin device when there is no limiting portion as in the case of Figure 81A and when there is a porous limiting portion as in the case of Figure 81B. The device without any limiting portion could be filled with approximately 87 million cells, and the device with a porous limiting portion could be filled with approximately 80 million cells. This indicates that the porous limiting portion can suppress membrane expansion and curvature during device filling. By using a spacer with a target distance between the porous limiting portions, the limiting portion can be further adjusted.

[0204] Example 31 In Vivo Transplantation into a Miniature Pig Model In this example, the transplantation study of mini-pigs into a human-sized ultra-thin device will be described. Ten mini-pigs were transplanted with an empty ultra-thin device, or an ultra-thin device filled with SC pancreatic islet cells (SEM-01) or porcine pancreatic islet cells at the preperitoneal or subcutaneous site. Figure 83A shows an example of an ultra-thin device with a 2.6 mm dot pitch of a human single-module design transplanted into a mini-pig. Figure 83B shows a targeted preperitoneal or subcutaneous transplantation site located about 3 inches away from the midline of the mini-pig and avoiding the rib margin. Figure 83C shows a subcutaneous incision by Bovie electrocautery in preparation for device transplantation. Figure 83D shows a preperitoneal incision by an illuminated retractor in preparation for device transplantation. The incision at the transplantation site can be adapted to different transplantation approaches. Figure 84A shows an example of subcutaneous placement of the ultra-thin device. Figure 84B shows an example of preperitoneal placement of the ultra-thin device. Figure 85 shows an example of a mini-pig two weeks after subcutaneous (SQ) and preperitoneal (PP) transplantation of the ultra-thin device. The image shows no macroscopic evidence of inflammation around the transplantation site, and the animals also showed no signs of distress or pain. The present invention provides, for example, the following items. (Item 1) A cell-containing device, (a) a first membrane having a first surface including a plurality of channels and a plurality of second surfaces facing the first surface; and (b) a second membrane facing and attached to the plurality of second surfaces of the first membrane, wherein the first membrane and the second membrane form an enclosed compartment having a surface area to volume ratio of at least about 40 cm -1 and the enclosed compartment provides a volume for containing cells within the device, the cell-containing device. (Item 2) The device according to item 1, wherein the compartment includes a single continuous open space. (Item 3) The device according to item 1, wherein the volume is about 8 uL to about 1,000 uL. (Item 4) ​The device according to item 1, wherein at least one of the length and the width is from about 0.25 cm to about 3 cm. (Item 5) The device according to item 1, wherein the thickness is at least about 300 μm. (Item 6) The device according to item 1, wherein the plurality of channels are substantially perpendicular to the first membrane. (Item 7) The device according to item 1, wherein the plurality of channels are arranged in a linear array. (Item 8) The device according to item 1, wherein the plurality of channels are arranged in a polar array. (Item 9) The device according to item 1, wherein the plurality of channels have an average diameter of from about 400 μm to about 3,000 μm. (Item 10) The device according to item 9, wherein the diameter is measured at the narrowest point within the plurality of channels. (Item 11) The device according to item 1, wherein the centers of each of the plurality of channels are separated by a distance of from about 75 μm to about 500 μm from the center of another channel. (Item 12) The device according to item 1, wherein the channels have a height-to-diameter ratio of at least about 0.2. (Item 13) The device according to item 1, wherein the device has a number of channels per area along a cross-section that is greater than about 50 / cm 2 The device according to item 1. (Item 14) The device according to item 1, wherein at least one of the first membrane and the second membrane includes a plurality of nodes interconnected by a plurality of small fibers. (Item 15) The device according to item 1, wherein at least one of the first membrane and the second membrane includes PVDF, PTFE, ePTFE, PCL, PE / PES, PP, PS, PMMA, PLGA, PLLA, or any combination thereof. (Item 16) The device according to item 1, further comprising an opening passing through the first membrane and the second membrane in the channel. (Item 17) The device according to item 16, wherein the opening has a maximum concentricity with respect to the channel and is 25% of the diameter of the channel. (Item 18) The device according to item 1, further comprising a frame configured to receive the device. (Item 19) The device according to item 18, wherein the frame is configured to receive a plurality of cell-containing devices. (Item 20) The device according to item 18, wherein the frame includes a flexible mechanism configured to prevent buckling of the cell-containing device. (Item 21) The device according to item 1, further comprising a cell population. (Item 22) The device according to item 21, wherein the cell population is an insulin-secreting population. (Item 23) The device according to item 21, wherein the cell population is stem cell-derived cells capable of glucose-stimulated insulin secretion (GSIS). (Item 24) The device according to item 1, further comprising a coating containing a hydrophilic polymer. (Item 25) The insulin diffusion coefficient is about 2x10-6 cm 2 / s to about 1x10-5 cm 2 / s. The device according to item 1. (Item 26) The device according to item 1, wherein the maximum oxygen diffusion distance is less than about 150 μm. (Item 27) The device according to item 1, wherein the first membrane and the second membrane are fused with a fusion detachment force of at least about 0.4 N. (Item 28) The device according to item 1, wherein at least one of the first membrane and the second membrane is semi-permeable. (Item 29) The device according to item 28, wherein the semi-permeability of the first membrane, the second membrane, or both is configured to protect the cells from immune attack. (Item 30) The device according to item 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. (Item 31) The device according to item 1, wherein at least one of the first membrane and the second membrane is configured to enable angiogenesis of the cells within the device. (Item 32) The composition according to item 31, wherein at least one of the first membrane and the second membrane is configured to enable angiogenesis of the cells within the device in the absence of immunosuppressive therapy. (Item 33) A cell-containing device, (a) a first membrane having a first surface including a plurality of channels and a plurality of second surfaces facing the first surface; and (b) a second membrane facing and attached to the plurality of second surfaces of the first membrane, wherein the first membrane and the second membrane form an enclosed compartment, the enclosed compartment provides a volume for accommodating 1 million to 1 billion insulin-producing cells within the device, and the membrane enables diffusion of insulin from the device while retaining the insulin-producing cells within the device, the cell-containing device. (Item 34) A composition comprising insulin-producing cells and a device for accommodating the insulin-producing cells, wherein when the device is transplanted into an individual, the device releases insulin while retaining the insulin-producing cells within the device and facilitates tissue angiogenesis within and around the device. (Item 35) The composition according to item 34, wherein the individual is not administered an immunosuppressant during the transplantation or angiogenesis of the device. (Item 36) The composition according to item 34, comprising 1 million to 1 billion insulin-producing cells. (Item 37) The composition according to item 34, wherein the device has a thickness of at least about 300 μm. (Item 38) The composition according to item 34, wherein the device comprises a membrane including a plurality of nodes interconnected by a plurality of small fibers. (Item 39) A method for manufacturing a cell-containing device, comprising: (a) providing a first membrane having a first surface and an opposing second surface; (b) forming a plurality of channels in the first surface of the first membrane; (c) fusing a second membrane to the second surface of the first membrane to form a compartment for containing cells between the second surface of the first membrane and the second membrane. (Item 40) Forming a plurality of channels in the first membrane comprises: (a) heating the first membrane at a predetermined time, a predetermined pressure, and a predetermined temperature; (b) shaping the plurality of channels using a mold, the method according to item 39. (Item 41) The method according to item 40, wherein fusing the second membrane to the first membrane is performed within the mold. (Item 42) The method according to item 41, wherein the mold includes a positive mold. (Item 43) The method according to item 41, wherein the mold includes a negative mold. (Item 44) The predetermined temperature is from about 100 degrees Celsius (°C) to about 600 °C, the method according to item 40. (Item 45) The method according to item 40, wherein the predetermined pressure is from about 2 pounds per square inch (psi) to about 140 psi. (Item 46) The method according to item 40, wherein the predetermined time is from about 3 minutes to about 30 minutes. (Item 47) The method according to item 40, wherein the predetermined pressure is about 3.5 psi and the predetermined temperature is about 370 °C. (Item 48) Forming a plurality of channels in the first membrane and fusing the second membrane to the first membrane comprises: (a) disposing the first membrane and the second membrane within a frame, wherein the first membrane and the second membrane are substantially parallel, substantially aligned, and separated by a gap distance; and (b) using a fusion tool to strike one or more points on the first membrane, wherein the fusion tool is heated to a set fusion temperature and contacts the membrane for a set fusion time between each strike, the method according to item 39. (Item 49) The method according to item 48, wherein striking the first membrane penetrates the first membrane, the second membrane, or both, and fuses a portion of the first membrane to the second membrane. (Item 50) The method according to item 48, wherein the frame surrounds at least a portion of the outer edges of the first membrane and the second membrane. (Item 51) The method according to item 48, wherein the gap distance is from about 300 μm to about 1,200 μm. (Item 52) The fusion tool has a strike contact area of at least about 0.07 mm 2 The method according to item 48. (Item 53) The method according to item 48, wherein dotting one or more points on the first membrane using a fusion tool includes dotting each of the one or more points up to about 16 times. (Item 54) The method according to item 53, wherein dotting one or more points on the first membrane using a fusion tool includes dotting each of the one or more points 1 to 6 times. (Item 55) The method according to item 53, wherein the set fusion temperature is from about 250°C to about 1,600°C. (Item 56) The method according to item 53, wherein the set fusion time is less than about 1 second. (Item 57) The method according to item 39, wherein at least one of the first membrane and the second membrane is substantially flat. (Item 58) The method according to item 39, further comprising embossing the first membrane before forming the plurality of channels in the first membrane. (Item 59) The method according to item 39, further comprising laser ablating a portion of the first membrane and the second membrane within the plurality of channels. (Item 60) The method according to item 59, wherein the fusion portion between the first membrane and the second membrane is removed by the laser ablation to form an opening. (Item 61) The method according to item 60, wherein the opening has a concentricity with respect to the channel of up to 25% of the diameter of the channel. (Item 62) The method according to item 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. (Item 63) The method according to item 39, further comprising coating the device with a hydrophilic polymer. (Item 64) The method according to item 39 of sintering the first membrane. (Item 65) The method according to item 39 of not sintering the second membrane. (Item 66) The method according to item 39, wherein the second membrane and the first membrane are fused with a fusion detachment force of at least about 0.2 N. (Item 67) A method comprising: a) contacting a tissue of a subject with diabetes or prediabetes with a device comprising a population of insulin-secreting cells, the device comprising: (i) a first membrane having a first surface including a plurality of channels and a plurality of second surfaces facing the first surface; and (ii) a second membrane facing and attached to the plurality of second surfaces of the first membrane, wherein the first membrane and the second membrane form an enclosed compartment having a surface area to volume ratio of at least about 40 cm -1 and the enclosed compartment provides a volume for containing cells within the device, the contacting; b) releasing insulin from the population of insulin-secreting cells in response to an increase in blood glucose level in the subject with diabetes, the increased glucose level being higher than the blood glucose level in a non-diabetic subject, the releasing. A method as described in any one of items 67, wherein the population of insulin-secreting cells releases an amount of insulin sufficient to lower the blood glucose level in the subject with diabetes or prediabetes. (Item 68) The method according to item 68, wherein releasing insulin stops when the blood glucose level in the subject with diabetes drops to a normal level. (Item 69) The method according to item 68, wherein releasing insulin stops when the blood glucose level in the subject with diabetes drops to a normal level. (Item 70) Releasing insulin, the method according to item 69 which resumes when the insulin-secreting cell population is re-exposed to an increase in blood glucose level in the diabetic subject. (Item 71) The method according to item 70, wherein the insulin-secreting cell population is a stem cell-derived cell population. (Item 72) The method according to item 71, wherein the insulin-secreting cell population is capable of glucose-stimulated insulin secretion (GSIS). (Item 73) The method according to item 68, wherein at least one of the first membrane and the second membrane is semipermeable. (Item 74) The method according to item 73, wherein the semipermeability of the first membrane, the second membrane, or both is configured to protect the cells from immune attack. (Item 75) The method according to item 74, wherein 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. (Item 76) The method according to item 68, wherein at least one of the first membrane and the second membrane is configured to enable angiogenesis of the cells in the device. (Item 77) The method according to item 76, wherein at least one of the first membrane and the second membrane is configured to enable angiogenesis of the cells in the device in the absence of immunosuppressive therapy.

Claims

1. A cell containment device, a first membrane, a second membrane disposed on the first membrane and attached to the first membrane, and a coating disposed on the first membrane and the second membrane comprising, wherein the first membrane and the second membrane form an enclosed compartment configured to contain a cell population within the device, the coating comprises a hydrophilic polymer, the coating comprises tetra(ethylene glycol) diacrylate (TEGDA) and hydroxypropyl acrylate (HPA), and the coating is crosslinked using an ammonium persulfate initiator, the first membrane and the second membrane comprise a porous biocompatible polymer, a cell containment device.

2. The device according to claim 1, further comprising a frame extending at least partially along the outer edges of the first membrane and the second membrane.

3. The device according to claim 1, wherein at least one of the first membrane and the second membrane comprises polyvinylidene fluoride (PVDF), polytetrafluoroethylene (PTFE), expanded polytetrafluoroethylene (ePTFE), polycaprolactone (PCL), polyethylene (PE), polyethersulfone (PES), polypropylene (PP), polystyrene (PS), poly(methyl methacrylate) (PMMA), poly(lactic-co-glycolic acid) (PLGA), poly(l-lactic acid) (PLLA), or any combination thereof.

4. The device according to claim 3, wherein at least one of the first membrane and the second membrane comprises ePTFE.

5. The device according to claim 1, wherein the volume of the compartment is 8 μL to 1,000 μL.

6. The device according to claim 1, wherein the device has a thickness of 10 μm to 500 μm.

7. The device according to claim 1, wherein the device has a maximum oxygen diffusion distance of less than 150 μm.

8. The device according to claim 1, further comprising a plurality of channels formed between the first membrane and the second membrane.

9. The device according to claim 8, wherein the plurality of channels have an average diameter of 400 μm to 3,000 μm.

10. The device has a number of channels per area along a cross-section greater than 50 / cm ２ The device according to claim 8, having a number of channels per area along a cross-section greater than 50 / cm

11. The device according to claim 1, further comprising the cell population disposed within the enclosed compartment.

12. The device according to claim 11, wherein the cell population is an insulin-secreting population.

13. The device according to claim 11, wherein the cell population is capable of glucose-stimulated insulin secretion (GSIS).

14. The first membrane and / or the second membrane has an insulin diffusion coefficient of 2×10^−6 cm ２ / s to 1×10^−5 cm ２ / s. The device according to claim 1

15. The device according to claim 1, wherein at least one of the first membrane and the second membrane is semi-permeable, and the semi-permeability of at least one of the first membrane and the second membrane is configured to protect the cell-containing device from immune attack and allow diffusion of a therapeutic compound.

16. The device according to claim 1, wherein at least one of the first membrane and the second membrane is sintered.

17. A method of manufacturing a cell-containing device, comprising: crosslinking a coating containing a hydrophilic polymer on a first membrane and a second membrane, the coating containing tetra(ethylene glycol) diacrylate (TEGDA) and hydroxypropyl acrylate (HPA), and the coating being crosslinked using an ammonium persulfate initiator; and fusing the second membrane to the first membrane to form an enclosed compartment configured to contain a cell population. Including wherein the first membrane and the second membrane comprise a porous biocompatible polymer. Method.

18. The method according to claim 17, wherein the crosslinking is performed before the fusing.

19. The method according to claim 17, wherein the crosslinking is performed after the fusing.

20. The method according to claim 17, further comprising forming a plurality of channels between the first membrane and the second membrane.

21. The method according to claim 17, wherein at least one of the first membrane and the second membrane comprises polyvinylidene fluoride (PVDF), polytetrafluoroethylene (PTFE), expanded polytetrafluoroethylene (ePTFE), polycaprolactone (PCL), polyethylene (PE), polyethersulfone (PES), polypropylene (PP), polystyrene (PS), poly(methyl methacrylate) (PMMA), poly(lactic-co-glycolic acid) (PLGA), poly(l-lactic acid) (PLLA), or any combination thereof.

22. The method according to claim 21, wherein at least one of the first membrane and the second membrane comprises ePTFE.

23. The method according to claim 17, wherein the volume of the compartment is from 8 μL to 1,000 μL.

24. The method according to claim 17, wherein the device has a thickness of from 10 μm to 500 μm.

25. The method according to claim 17, wherein the device has a maximum oxygen diffusion distance of less than 150 μm.

26. The method according to claim 17, further comprising filling the compartment with the cell population.

27. The method according to claim 26, wherein the cell population is an insulin-secreting population.

28. The method according to claim 26, wherein the cell population is capable of glucose-stimulated insulin secretion (GSIS).

29. The first membrane and / or the second membrane has an insulin diffusion coefficient of 2×10^−6 cm ２ / s to 1×10^−5 cm ２ / s, the method according to claim 17.

30. The method according to claim 17, wherein at least one of the first membrane and the second membrane is semipermeable, and the semipermeability of at least one of the first membrane and the second membrane is configured to protect the cell-containing device from immune attack and allow diffusion of a therapeutic compound.

31. The method according to claim 17, wherein at least one of the first membrane and the second membrane is sintered.

32. The method according to claim 17, further comprising sintering at least one of the first membrane and the second membrane.

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