Assembly of a three-dimensional large-capacity cell-encapsulating device

Abstract

To provide implantable 3-dimensional large capacity device assemblies, specifically, large capacity device assemblies for encapsulating pancreatic progenitor cells for treatment of diabetes.SOLUTION: Provided is an encapsulation device, preferably a cell encapsulation device, preferably a macrophage encapsulation device, preferably a large capacity device assembly, preferably a cell encapsulation device assembly with any size which consists of a device comprising at least one, two, three, four, five, six, seven, eight, nine, ten, or more cell chambers.SELECTED DRAWING: Figure 3

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application is a continuation of U.S. Provisional Application No. 61 / 774,443, filed March 7, 2013. The benefit of this provisional application is claimed, and this provisional application is incorporated herein in its entirety for all purposes. do.

[0002] FIELD OF THE INVENTION The field of the invention relates to medical devices and cell therapy. One embodiment relates to high-capacity encapsulation of cells by a semipermeable implantable device. Summary of the Invention

[0003] The embodiments described herein include a method for implanting a population of live cells into a mammalian recipient, comprising: Cell encapsulation assembly, large capacity device assembly, or 3D large capacity device Regarding assembly.

[0004] In one embodiment, the large capacity device assembly comprises at least two cell chambers. , and the fold and unfold configurations It contains at least two configurations of the field configuration. The footprint of the folded configuration is smaller than that of the unfolded configuration.

[0005] As used herein, footprint refers to the projection of a device onto a two-dimensional plane on an anatomical site. In one embodiment, the method comprises at least two chambers for encapsulating living cells. A large capacity device assembly for implantation into a mammalian recipient, comprising: a first seal at the outer periphery of the battery, forming a second seal to form an enclosed assembly; a seal portion of the cell encapsulation assembly and a seal portion of the cell chamber that forms the inner periphery of the cell encapsulation assembly; and at least a second sealing portion. The breaker may have a third or fourth seal that further separates each of the cell chambers. , i.e., a septum seal portion.

[0006] In one embodiment, the large capacity device assembly is three-dimensional and includes a Roman sheath. Shape of the tube, U-shape, scallop, fin shape, flat tube, coil, fan, radiator ator, or encapsulating a therapeutically effective dose of cells while limiting the footprint of the assembly. It is possible to take any other three-dimensional shape.

[0007] In one embodiment, the large capacity device assembly is inserted into the body of a recipient and its shape It is a three-dimensional assembly that can maintain its shape, form and position.

[0008] In one embodiment, the cell chamber of the cell encapsulation assembly comprises a lumen-like cell matrix. The cells in the chamber, particularly the cells in the core or center of the chamber, The lumenal matrix provides improved oxygen and nutrient exchange with the alveoli. The material may be a silicone material such as (but not limited to) silicone foam or silicone fiber. In another embodiment, the lubricant may comprise an elastomer matrix, including a cone elastomer. The membrane matrix acts as a conduit, allowing the flow of oxygen and nutrients to the enclosed cells. thereby enhancing short-term and long-term cell survival after implantation. is any biostable agent that promotes

[0009] In one embodiment, the large capacity device assembly comprises at least two cell chambers. , and the fold and unfold configurations It contains at least two configurations of the field configuration. The footprint of the configuration is smaller than the unfolded configuration, but the surface area is the same. is.

[0010] In one embodiment, the large capacity device assembly, when implanted in a mammalian recipient, A fold configuration to flatten or unfold, In this embodiment, the incision site is small, but the implantation site contains at least two cell chambers. Once inserted, the assembly flattens and spreads, reducing its extrusion from the recipient. Minimize insertion and maximize insertion.

[0011] In one embodiment, the large capacity device assembly is in a first unfolded configuration. Simulation, Second Fold Configuration, and Fold Configuration Includes a third implant configuration that is flatter than the standard implant configuration.

[0012] In one embodiment, the large capacity device assembly is a flat assembly with the same footprint. A fold configuration with at least twice as many viable cells as the It contains at least two cell chambers.

[0013] Preferred features and embodiments of the present invention are as follows:

[0014] In a preferred embodiment, the assembly comprises more than one cell for encapsulating a living cell. In a preferred embodiment, the assembly includes a small chamber for encapsulating living cells. It contains at least two cell chambers.

[0015] In a preferred embodiment, the assembly comprises a cell-free region. The assembly includes a cell-free region along the longitudinal axis that separates the cell chambers. In embodiments, the assembly includes a cell-free region that bends to form a fold. In a preferred embodiment, the folds increase the footprint of the assembly by a factor of 10 compared to the footprint of the assembly without the folds. Reduced compared to Assembly.

[0016] In a preferred embodiment, the assembly may be performed with or without folding. The cell volume capacity remains substantially the same even when the cell volume is not increased.

[0017] In a preferred embodiment, the assembly includes a semi-permeable membrane.

[0018] In preferred embodiments, the assembly is comprised of two, three, four, five, six, seven, eight, or , or more cell chambers.

[0019] In a preferred embodiment, the assembly includes at least one loading port. In a preferred embodiment, the assembly includes two loading ports.

[0020] In a preferred embodiment, the living cells are definitive endoderm lineage cells. In a preferred embodiment, the living cells are , human PDX1 (pancreatic and duodenal homeobox In a preferred embodiment, the viable cells are human endocrine disrupting cells. In a preferred embodiment, the living cells are human immature beta cells. In a preferred embodiment, the cells are dispersed within the chamber.

[0021] In a preferred embodiment, the cell chamber disperses live cells and Having a matrix with multiple interconnected cavities or pores that improve oxygen distribution In a preferred embodiment, the interconnected cavities have different cavity sizes. In the matrix, polydimethylsiloxane (PDMS) monoacrylate, and polydimethylsiloxane monomethacrylate. In one embodiment, the matrix is ​​a silicone elastomer.

[0022] In a preferred embodiment, the cell chambers are parallel to one another. In this embodiment, the cell chambers are spaced apart by about 20 degrees. The members are spaced about 40 degrees apart. In a preferred embodiment, the chamber is a cell chamber. Includes bulkhead seal within bar. [Brief explanation of the drawings]

[0023] [Figure 1] Figure 1 is a graph showing beta cell mass and relative islet equivalent (IEQ) per kg of body weight (BW). The graph also notes that while the onset of diabetes occurs at approximately 10-20% beta cell mass, serum C-peptide is not discernible in patients with beta cell mass below 10%, and that although the therapeutic index range is wide, approximately 200,000 IEQ is a potentially effective dose delivered by encapsulated PEC grafts.

[0024] [Figure 2]FIG. 2 is a graph correlating human islet IEQ and C-peptide content with C-peptide derived from mature encapsulated pancreatic endoderm cell (PEC) grafts.

[0025] [Figure 3A-B] Figure 3 is a perspective view of one embodiment of a three-dimensional large capacity device assembly. Figure 3B is a cross-sectional view of Figure 3A showing the three-dimensional nature of the device assembly folded at an angle such that the cell chambers are substantially parallel to one another.

[0026] [Figure 4A-B] Figure 4 shows photographs of one embodiment of a three-dimensional large capacity device assembly: Figure 4A shows a flat, planar, eight-cell chamber device, and Figure 4B shows the same device of Figure 4A folded so that the cell chambers are substantially parallel to each other.

[0027] [Figure 5A-B] Figure 5 shows photographs of cell encapsulation devices: Figure 5A shows a three-dimensional large capacity device assembly with dual ports, compared to the small capacity planar device shown in Figure 5B.

[0028] [Figure 6A-C] Figure 6 shows a perspective view of one embodiment of a three-dimensional, large-capacity device assembly without ports: Figure 6A shows the three-dimensional nature of the device assembly with folds at angles such that the cell chambers are substantially parallel to one another or spaced apart at 0 degrees, Figure 6B shows a top view of the device, and Figure 6C shows a cross-section of the device.

[0029] [Figure 7A-B]Figure 7 shows a perspective view of one embodiment of a three-dimensional, high-capacity device assembly with ports: Figure 7A shows the three-dimensional nature of the device assembly, and Figure 7B shows a cross-section of the device with ports, with each cell chamber and port spaced approximately 20 degrees apart.

[0030] [Figure 8A-B] Figure 8 shows a perspective view of one embodiment of a three-dimensional, high-capacity device assembly with ports: Figure 8A shows the three-dimensional nature of the device assembly, and Figure 8B shows a cross-section of the device with ports, with each cell chamber and port spaced approximately 40 degrees apart.

[0031] [Figure 9A-C] Figure 9 shows a perspective view of one embodiment of a three-dimensional, large-capacity device assembly without ports ("Roman shade" style). Figure 9A shows the three-dimensional nature of the device assembly, Figure 9B shows a top view of the device, and Figure 9C shows a cross-section of the device without ports. The cell chambers are parallel to each other but at an angle.

[0032] [Figure 10A-C] Figure 10 shows a perspective view of one embodiment of a three-dimensional large capacity device assembly in which the chamber is a continuous tube: Figure 10A shows a cross section of the tubular device of Figure 10B, cut in half to show details of the serpentine cell chamber, Figure 10B shows a flat sheet of the tubular device with openings at both ends, and Figure 10C shows a top view of the tubular device.

[0033] [Figure 11A-C]Figure 11 shows a perspective view of one embodiment of a three-dimensional large capacity device assembly: Figure 11A shows a three-dimensional large capacity device, Figure 11B shows a top view of the device, and Figure 11C shows a cross-section of the device with the cell chambers parallel to each other and attached to a base on one side.

[0034] [Figure 12A-C] Figure 12 shows a perspective view of one embodiment of a ("shutter" type) three-dimensional large capacity device assembly: Figure 12A shows a three-dimensional large capacity device, Figure 12B shows a top view of the device, and Figure 12C shows a cross-section of the device with parallel cell chambers interconnected to the base.

[0035] [Figure 13A-B] Figure 13 shows top perspective views of two embodiments of cell-encapsulating large-capacity device assemblies containing eight cell chambers with one port (Figure 13A) or two ports (Figure 13B) before being formed or folded into a three-dimensional large-capacity device assembly.

[0036] [Figure 14A-B] Figure 14 shows top perspective views of two embodiments of a cell-encapsulating large-capacity device assembly (Figure 14A) containing 16 cell chambers with one port; and modular fabrication of device assemblies with one, two, three or more cell chambers with one port.

[0037] [Figure 15] FIG. 15 is a perspective view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port per cell chamber.

[0038] [Figure 16]FIG. 16 is a rear elevation view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port per cell chamber.

[0039] [Figure 17] FIG. 17 is a front elevation view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port per cell chamber.

[0040] [Figure 18] FIG. 18 is a top view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port per cell chamber.

[0041] [Figure 19] FIG. 19 is a bottom view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port per cell chamber.

[0042] [Figure 20] FIG. 20 is a right elevation view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers, each with a port (circle), where the cell chambers are parallel to each other.

[0043] [Figure 21] Figure 21 is a left elevation view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers, each with a port (circle), where the cell chambers are parallel to each other.

[0044] [Figure 22]FIG. 22 is a perspective view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port per cell chamber.

[0045] [Figure 23] FIG. 23 is a rear elevation view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port per cell chamber.

[0046] [Figure 24] FIG. 24 is a front elevation view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port per cell chamber.

[0047] [Figure 25] FIG. 25 is a top view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port per cell chamber.

[0048] [Figure 26] FIG. 26 is a bottom view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port per cell chamber.

[0049] [Figure 27] Figure 27 is a right elevation view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers, each with a port (circle), where the cell chambers are parallel to each other.

[0050] [Figure 28]Figure 28 is a left elevation view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers, each with a port (circle), where the cell chambers are parallel to each other.

[0051] [Figure 29] FIG. 29 is a perspective view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber in the shape and form of a tube with a port at each end.

[0052] [Figure 30] FIG. 30 is a rear elevation view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber in the shape and form of a tube with a port at each end.

[0053] [Figure 31] FIG. 31 is a front elevation view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber in the shape and form of a tube with a port at each end.

[0054] [Figure 32] FIG. 31 is a top view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber in the shape and form of a tube with a port at each end.

[0055] [Figure 33] FIG. 33 is a bottom view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber in the shape and form of a tube with a port at each end.

[0056] [Figure 34]FIG. 34 is a right elevation view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber in the shape and form of a tube with a port at each end.

[0057] [Figure 35] FIG. 35 is a left elevation view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber in the shape and form of a tube with a port at each end.

[0058] [Figure 36] FIG. 36 is a perspective view of a three-dimensional large capacity cell encapsulation device or assembly constructed from a single modular unit with a cell chamber on each side.

[0059] [Figure 37] FIG. 37 is a rear elevation view of a three-dimensional large capacity cell encapsulation device or assembly constructed from a single modular unit with a cell chamber on each side.

[0060] [Figure 38] Figure 38 is a front elevation view of a three-dimensional large capacity cell encapsulation device or assembly constructed from modular cell chamber units; the figure shows eight such units assembled, although the device may have more or fewer units assembled.

[0061] [Figure 39] FIG. 39 is a top view of a three-dimensional large capacity cell encapsulation device or assembly constructed from a single modular unit with a cell chamber on each side.

[0062] [Figure 40]FIG. 40 is a bottom view of a three-dimensional large capacity cell encapsulation device or assembly constructed from a single modular unit with a cell chamber on each side.

[0063] [Figure 41] FIG. 41 is a right elevation view of a three-dimensional large capacity cell encapsulation device or assembly constructed from a single modular unit with a cell chamber on each side.

[0064] [Figure 42] FIG. 42 is a left elevation view of a three-dimensional large capacity cell encapsulation device or assembly constructed from a single modular unit with a cell chamber on each side.

[0065] [Figure 43] FIG. 43 is a perspective view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port.

[0066] [Figure 44] FIG. 44 is a rear elevation view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port.

[0067] [Figure 45] FIG. 45 is a front elevation view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port.

[0068] [Figure 46] FIG. 46 is a top view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port.

[0069] [Figure 47] FIG. 47 is a bottom view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port.

[0070] [Figure 48] Figure 48 is a right elevation view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port (circle), with the cell chambers facing parallel to each other.

[0071] [Figure 49] Figure 49 is a left elevation view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port (circle), with the cell chambers facing parallel to each other.

[0072] [Figure 50] FIG. 50 is a perspective view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber and a single port.

[0073] [Figure 51] FIG. 51 is a rear elevation view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber and a single port.

[0074] [Figure 52] FIG. 52 is a front elevation view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber and a single port.

[0075] [Figure 53] FIG. 53 is a top view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber and a single port.

[0076] [Figure 54] FIG. 54 is a bottom view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber and a single port.

[0077] [Figure 55] FIG. 55 is a right elevation view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber and a single port.

[0078] [Figure 56] FIG. 56 is a left elevation view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber and a single port.

[0079] [Figure 57] FIG. 57 is a perspective view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port, the assembly closely resembling a plantation shutter design.

[0080] [Figure 58] FIG. 58 is a rear elevation view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port.

[0081] [Figure 59] FIG. 59 is a front elevation view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port.

[0082] [Figure 60]FIG. 60 is a top view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port.

[0083] [Figure 61] FIG. 61 is a bottom view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port.

[0084] [Figure 62] FIG. 62 is a right elevation view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port (circle), the assembly closely resembling a plantation shutter design.

[0085] [Figure 63] FIG. 63 is a left elevation view of a three-dimensional large capacity cell encapsulation device or assembly with multiple cell chambers and a single port, the assembly closely resembling a plantation shutter design.

[0086] [Figure 64] FIG. 64 is a perspective view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber and a single port, the assembly closely resembling a plantation shutter design.

[0087] [Figure 65] FIG. 65 is a rear elevation view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber and a single port.

[0088] [Figure 66]FIG. 66 is a front elevation view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber and a single port.

[0089] [Figure 67] FIG. 67 is a top view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber and a single port.

[0090] [Figure 68] FIG. 68 is a bottom view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber and a single port.

[0091] [Figure 69] FIG. 69 is a right elevation view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber and a single port, the assembly closely resembling a plantation shutter design.

[0092] [Figure 70] FIG. 70 is a left elevation view of a three-dimensional large capacity cell encapsulation device or assembly with a single cell chamber and a single port, the assembly closely resembling a plantation shutter design.

[0093] [Figure 71A-C] Figure 71 shows photographic images of a silicone-based hollow fiber tube (Figures 71A-B) and a silicone-based elastomeric foam (Figure 71C) woven to form a mat for use within the cell chamber of a cell encapsulation device assembly.

[0094] [Figure 72]Figure 72 is a graph showing the concentration of human C-peptide in the serum of implanted mice for six experimental and six control animals. In vivo glucose level response function was analyzed 13 weeks after implantation or transplantation during fasting and 30 and 60 minutes after intraperitoneal glucose administration. All animals received encapsulated PEC implants (Encaptra® EN20 or EN20; ViaCyte, San Diego, CA) with or without a silicone hollow fiber lumenal matrix.

[0095] [Figure 73A-B] Figure 73 shows photographic images of histology sections of explanted PEC grafts with silicone hollow fibers. The hollow fibers are the white circular structures between the semipermeable membranes of the device. The sections were stained with standard hematoxylin and eosin stain (Figure 73A) and an anti-insulin antibody (Figure 73B), which stains those cells expressing insulin brown.

[0096] [Figure 74] Figures 74A-C are ultrasound images showing a prototype of the 3D cell encapsulation device assembly of the EN250 device (Figure 74A) implanted in a human (recently deceased) cadaver and imaged by ultrasound. The U-shaped prototype of the EN250 device shown can be viewed before, during, and after application of a compressive load to the cadaver (Figures 74B and C).

[0097] [Fig. 75A-B] FIG. 75 shows ultrasound images of the wetted empty device and the filled EN250 and EN20 devices. DETAILED DESCRIPTION OF THE INVENTION

[0098] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Unless otherwise stated, terms used herein are used in accordance with conventional usage by those skilled in the art. It is to be understood that various patent and non-patent publications are referenced throughout this application. The entire disclosures of these publications and the references cited therein are set forth in their entirety. In order to more fully describe the state of the art to which the patents pertain, they are incorporated herein by reference. is incorporated by reference in its entirety into this application.

[0099] For purposes of this specification and the appended claims, unless otherwise indicated, All numbers, ingredients, and amounts of ingredients used in this specification and claims are intended to be used interchangeably. Percentages or ratios, reaction conditions, and other numerical values ​​are also expressly provided by the term "about" in all cases. It should be understood that the term is modified by the term.

[0100] Accordingly, unless indicated to the contrary, the following specification and accompanying The numerical parameters set forth in the claims may vary depending upon the desired properties sought to be obtained. This is an approximation. It is not intended as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, but at a minimum, To the extent possible, each numerical parameter should at least be within the bounds of the usual standard given the number of reported significant digits. shall be interpreted by applying rounding methods.

[0101] In one embodiment, a biocompatible implantable device is provided. Inlet devices are described in U.S. Patents 6,773,458; 6,156,305; No. 060,640; No. 5,964,804; No. 5,964,261; No. 5,8 No. 82,354; No. 5,807,406; No. 5,800,529; No. 5,78 No. 2,912; No. 5,741,330; No. 5,733,336; No. 5,713 , No. 888; No. 5,653,756; No. 5,593,440; No. 5,569, No. 462; No. 5,549,675; No. 5,545,223; No. 5,453,2 No. 78; No. 5,421,923; No. 5,344,454; No. 5,314,47 No. 1; No. 5,324,518; No. 5,219,361; No. 5,100,392 and 5,011,494, all of which are incorporated herein by reference in their entirety. It has been transferred to ter.

[0102] Other suitable embodiments described herein are in accordance with at least one patent application Ser. No. 09 / 109,523, issued Jul. 3, 2012. No. 8,211,699, METHODS FOR CULTURING PLURIPOTENT STEM CELLS IN SUSPENSION US ING ERBB3 LIGANDS; No. 7,95 issued on July 26, 2011 No. 8,585, PREPRIMITIVE STREAK AND MESENDODE RM CELLS; issued on March 31, 2009 and July 10, 2012, respectively. Nos. 7,510,876 and 8,216,836, DEFINITIVE ENDODERM; Issued June 2, 2009, No. 7,541,185, M ETHODS FOR IDENTIFYING FACTORS FOR DIFFE RENTIATING DEFINITIVE ENDODERM;December 1, 2009 No. 7,625,753, issued on the same day, EXPANSION OF DEFINIT IVE ENDODERM; No. 7,695,963 issued on April 13, 2010 No., METHODS FOR INCREASING DEFINITIVE ENDO DERM PRODUCTION; Issued on April 27, 2010, No. 7,704, No. 738, DEFINITIVE ENDODERM; issued August 9, 2011 No. 7,993,916, METHODS FOR INCREASING DEFI Native Endoderm Production; Issued August 30, 2011 No. 8,008,075, STEM CELL AGGREGATE SUSPENSION ENSION COMPOSITIONS AND METHODS OF DIFFE RENTIATION THEREOF; Issued on May 29, 2012, No. 8,1 No. 78,878, COMPOSITIONS AND METHODS FOR SEL F-RENEWAL AND DIFFERENTIATION IN HUMAN E MBRYONIC STEM CELLS; Issued on July 10, 2012, No. 8, No. 216,836, METHODS FOR IDENTIFYING FACTORS FOR DIFFERENTIATING DEFINITIVE ENDODERM ; exchanged on May 19, 2009, April 13, 2010, and August 9, 2011, respectively Nos. 7,534,608, 7,695,965 and 7,993, No. 920; No. 8,129,182 issued on March 6, 2012, ENDOCRI NE PRECURSOR CELLS,PANCREATIC HORMONE EXP RESSING CELLS AND METHODS OF PRODUCTION; No. 8,338,170, issued December 25, 2012, METHODS FO R PURIFYING ENDODERM AND PANCREATIC ENDO DERM CELLS DERIVED FROM HUMAN EMBRYONIC STEM CELLS; No. 8,334,138 issued December 18, 2012 , METHODS AND COMPOSITIONS FOR FEEDER-FRE E PLURIPOTENT STEM CELL MEDIA CONTAINING HUMAN SERUM; No. 8,278,106 issued on October 2, 2012 No., ENCAPSULATION OF PANCREATIC CELLS DERI VED FROM HUMAN PLURIPOTENT STEM CELLS;20 METHOD FOR PURIFYING END, issued on December 25, 2012 ODERM AND PANCREATIC ENDODERM CELLS DERI VED FROM HUMAN EMBRYONIC STEM CELLS(CYTH No. 8,338,170, entitled "ERA.063A" filed February 6, 2013; US Application No. 13 / 761,078, CELL COMPOSITIONS DERI VED FROM DEDIFFERENTIATED REPROGRAMMED C ELLS; U.S. Application No. 13 / 672,688, filed November 8, 2012, SC ALABLE PRIMATE PLURIPOTENT STEM CELL AGG REGATE SUSPENSION CULTURE AND DIFFERENTI ATION THEREOF; Design Patent Application No. 29 filed December 12, 2001 / 408,366; No. 29 / 408,368; and No. 29 / 408,370 and in US Patent Application No. 29 / 423,365, filed May 31, 2012, These are described in more detail in:

[0103] (definition) As used herein, "about" means that the number referred to as "about" is the recited number plus or minus that number. For example, "about" 100 cells means that the number of cells in the sample is between 1 and 10% of the number listed. Depending on the situation, this could mean 95-105 cells, or as few as 99-101 cells. Whenever it appears in this specification, it may refer to "1 to 20" or the like. Numeric ranges refer to each integer within a given range, e.g., "1 to 20 cells" refers to 1 cell, 2 cells, 3 cells, etc. means up to 20 cells, including 20 cells When "about" modifies a range expressed by numbers other than integers, the recited number plus or minus the expressed significant number. For example, approximately 1.50 to 2.50 mM means a minimum of 1 It can mean .35M, or up to 2.75M, and anything in between. , or any amount in increments of 0.01.

[0104] As used herein in the context of the composition of a cell population, "essentially" or "substantially" refers to The term "" means "mostly" or "mainly."

[0105] As used herein, the term "effective amount" of a compound or its equivalents refers to a feeder 1 month in the absence of cells and in the absence of serum or serum replacement A residual amount of defined medium sufficient to provide stabilization of differentiable cells in culture for over a period of time. This refers to the concentration of a compound in the presence of other components. This concentration can be easily determined by one skilled in the art. can be.

[0106] "Cells," "cell lines," "cell cultures," or "cell populations" or "populations of cells" The term "isolated" as used herein when referring to a living cell, cell line, Cell cultures, cell populations or cell populations in vitro for extended periods of time It refers to being substantially separated from the source of cells so that they can be cultured. The term "isolating" also refers to the isolation or separation of one or more cells from a group of two or more cells. physical selection of cells based on cell shape and / or expression of various markers; It can be used to refer to a physical choice that is selected based on

[0107] As used herein, the term "substantially" refers to a great extent or degree, e.g. For example, "substantially similar" in one context means something that is substantially similar to another method or to a greater extent than another method. However, the present invention is not limited to the above. The term "substantially free" as used in the Subpart B, e.g., "substantially free" or "Substantially free of contaminants" or "Substantially free of serum" or "Insulin or substantially free of insulin-like growth factors" or their equivalents are solutions, media , supplements, excipients, etc. are at least 98% or less free of serum, contaminants, or their equivalents. or at least 98.5%, or at least 99%, or at least 99.5%, or In one embodiment, a defined culture medium is and wherein the serum is not present or is 100% serum-free or substantially serum-free. Conversely, the term "substantially similar" as used herein means that the culture medium is not The term or its equivalents refer to compositions, processes, methods, solutions, media, supplements, excipients, etc., that are used in the present specification. or incorporated herein in its entirety. Compositions, steps, methods, solutions, media, supplements, excipients, etc. in the process or method being studied and at least 80%, at least 85%, at least 90%, at least 95%, and means at least 99% similar.

[0108] As used herein, cells suitable for transplantation are cells that can be used to treat metabolic disorders in vivo. a cell or population of cells that is sufficiently viable and / or functional to treat the For example, diabetes or one or more symptoms thereof may be associated with transplantation into a subject with diabetes. After implantation of suitable cells, symptoms can be improved or alleviated over a period of time. In embodiments, the cell or cell population suitable for transplantation is a pancreatic progenitor cell or population, or PDX1-positive pancreatic progenitor cells or populations, or endocrine progenitor cells or populations, or multi- Hormonal endocrine cells or single hormonal endocrine cells, and / or cells or Any combination of populations of cells, or PECs or further purified or enriched cells thereof. A cell or a group of cells.

[0109] Implantable high-capacity devices One embodiment described herein is an encapsulation device, preferably a cell encapsulation device, preferably a Preferably a macro cell encapsulation device, preferably a large capacity device assembly, preferably is at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more A cell encapsulation device assembly of any size, consisting of a device with one or more cell chambers. As used herein, the term "assembly" refers to a plurality of a cell encapsulation device consisting of a single or a plurality of cell chambers In one embodiment, the assembly includes at least one, two, three, four, Consists of 5, 6, 7, 8, 9, 10 or more cell chambers. In embodiments, the assembly may comprise any number of cell chambers (or modules). For example, a modular unit can consist of one , 2, 3, 4, 5, 6, 7, 8, 9, 10 or more cells It may consist of multiple members, depending on the number or dose of cells required to treat the disease. Therefore, the term "device" as used herein refers to any device that has been previously described. It may also refer to a single device consisting of one cell chamber, such as a cell chamber containing and a plurality of fine particles, such as the three-dimensional devices or device assemblies described herein. It can also refer to a single device consisting of a cell chamber. In this case, the terms device and assembly can be used interchangeably.

[0110] In one embodiment, the device or assembly has a total volume of about 20 μL, 50 μL, 1 00μL, 150μL, 200μL, 250μL, 300μL, 350μL, 400μL , 450μL, 500μL, 550μL, 600μL, 650μL, 700μL, 750 μL, 800μL, 850μL, 900μL, 950μL, 1000μL or more The total cell volume can be determined by the amount of one cell chamber that can hold the desired cell dose. It may consist of one device, which together have the desired cell dose, and any number or or a plurality of cell chambers, In one embodiment, the method previously described in U.S. Pat. No. 8,425,928 may be used. As described above, by creating one or more compartments within the cell chamber, Improve the device. Figures 3-70 show embodiments of the device or assembly. The device or assembly may be the device or assembly illustrated in FIGS. It is not intended to be limited to only the device or application. The assembly may include modifications based on the devices or assemblies described herein. It is possible, and this is considered routine in the art. , the device design depends on the type of bioactive agent and / or cell to be encapsulated, and , can be modified to meet the needs and functions of the study.

[0111] Such devices and / or assemblies can be implanted into mammals to produce a variety of Diseases and disorders can be treated. In a preferred embodiment, the device is Biocompatible materials capable of completely encapsulating bioactive drugs and / or cells within them. For example, such devices may be used to maintain oxygen and cell viability. Other molecules important for immune function can move through the semipermeable membrane, but cells of the immune system cannot pass through the pores. The therapeutically effective agent is placed in a semipermeable membrane having a pore size that is so small that the agent cannot penetrate or cross the membrane. Similarly, such devices may contain therapeutically effective amounts of bioactive agents, e.g. For example, they may contain angiogenic factors, growth factors, hormones, etc., and are secreted by cells. The composition may also contain bioactive agents, such as antibodies, proteins, hormones, etc.

[0112] The devices and / or assemblies described herein are suitable for the delivery of bioactive substances to biological Such devices can be employed to treat conditions requiring a continuous supply of The agent may also contain, for example, a homogeneous or heterogeneous mixture of bioactive agents and / or cells, or or bioartificial organs containing cells that produce one or more target bioactive substances. The bioactive agent and / or cells are contained within at least one semipermeable membrane. completely within at least one interior space or enclosed chamber bounded by Ideally, such a semipermeable membrane would encapsulate or close the enclosed space. The passage of biologically active substances (e.g., insulin, glucagon, pancreatic polypeptide, etc.) The active substance should be available to target cells both outside the device and within the patient's body. In a preferred embodiment, the semi-permeable membrane allows nutrients naturally present in the subject to pass through the membrane. This allows the cells to pass through and deliver essential nutrients to the encapsulated cells. Such semi-permeable membranes allow the patient's cells, more specifically immune system cells, to pass through the device and into the device. inhibit or prevent the passage of the ions into the device and harm the cells encapsulated within the device For example, in the case of diabetes, this approach may improve the recognition and function of the implanted cells by immune system cells. Glucose and oxygen are delivered in real time to the body on demand, preventing damage and breakdown. It may be possible to stimulate insulin-producing cells to release insulin. In certain embodiments, the semipermeable membrane inhibits implanted cells from escaping encapsulation.

[0113] A preferred device or assembly preferably comprises (but is not limited to): It may have certain characteristics, one or a combination of: i) The footprint of the device, e.g., the size of the device or assembly at the desired anatomical site. Large or high cell doses while simultaneously limiting the space occupied by the Containing a three-dimensional configuration that allows for delivery; ii) at the weld or where the device is sealed, or within the cell chamber The fold angle is 0 (or 180) to 90 degrees, preferably 0 to 50 degrees, and more preferably includes folds or bends or angles ranging from 0 to 40 degrees; iii) biocompatible materials that function under physiological conditions, including pH and temperature, e.g. Anisotropic materials (including but not limited to), polysulfone (PSF), nanofiber mats, polyimides , tetrafluoroethylene / polytetrafluoroethylene (PTFE: tetraflu oroethylene / polytetrafluoroethylene;also T eflon®), ePTFE (expanded polytetrafluoroethylene Ethylene: expanded polytetrafluoroethylene), poly Triacrylonitrile, polyethersulfone, acrylic resin, cellulose acetate, nitric acid cell Contains cellulose, polyamide, and hydroxypropyl methylcellulose (HPMC) membrane thing; iv) Harm or cause harm to bioactive agents and / or cells encapsulated within the device does not release toxic compounds that could damage them; v) promoting the secretion or release of bioactive agents or macromolecules across the device; iv) promoting rapid kinetics of macromolecular diffusion; vi) promoting the long-term stability of encapsulated cells; vii) promoting angiogenesis; viii) containing a chemically inert membrane or containment structure; ix) Providing stable mechanical properties; x) Maintaining structural / containment integrity (e.g., preventing unintended, toxic, or harmful drugs) preventing leakage of agents and / or cells); xi) be refillable and / or flushable; xii) mechanically extensible; xiii) Contains no ports or contains at least one, two, or more than two ports containing; xiv) immunoisolation of transplanted cells from recipient tissue; xv) ease of fabrication and manufacture; xvi) Ability to be sterilized; xvii) Capable of being manufactured in a modular fashion; xviii) be retrievable after implantation; xix) Ventilation during cell or therapeutic loading.

[0114] The embodiments of the encapsulation device described herein may include any of the above elements. To the extent that this is achievable, certain device sizes, shapes, designs, capacity, and and / or is not intended to be limited to the materials used to fabricate the encapsulation device. .

[0115] Encapsulation provides a protective barrier that prevents elements of the recipient's immune system from destroying the cells. This allows for the transfer of unmatched human tissue or animal tissue without recipient immunosuppression. Allows for the use of tissues and therefore an increased diversity of cell types that can be employed therapeutically. In addition, the implanted cells are held in place by a membrane, which prevents cell encapsulation. The introduction of ATP reduces the inherent risk of tumor formation that may otherwise be present in cell-based therapy. It also prevents sex.

[0116] The tissue or cells within the core of the device are then supported by a hydrogel or extracellular matrix. The components may also be immobilized on an immobilization matrix. , so as to further reduce the possibility of a necrotic core of cells in the center of the device. The cells may contain an insert that creates a "cell-free" zone in the center of the cell.

[0117] In a preferred embodiment, the device is immunoisolatory. Once implanted in a mammalian recipient, the recipient's immune system attacks cells within the core of the device. To be immunoisolating, the surrounding or peripheral area of ​​the device: (a) The encapsulated cells are exposed to the immune system of the recipient into whom the device or assembly is implanted. (b) to prevent harmful substances in the recipient's body from entering the core of the device; (c) sufficient physical protection to prevent adverse immunological contact between the isolated cells and the recipient's immune system; The thickness of this physical barrier can vary, but always On either side, sufficient pressure must be applied to prevent direct contact between cells and / or materials. The thickness of this region generally ranges from 5 to 200 microns, but is typically 10 A thickness of up to 100 microns is preferred, and a thickness of 20 to 75 microns is particularly preferred. The types of immune attacks that can be prevented or minimized by the use of phage, neutrophils, cell-mediated immune responses (e.g., natural killer cells and antibody-dependent T cell-mediated cytolysis (ADCC), as well as humoral responses (e.g., antibody-dependent This includes attacks by complement-mediated cytolysis.

[0118] The device maintains bioactivity and provides access for delivery of a product or function. Any configuration suitable for carrying out the process, e.g., cylindrical, rectangular, disc-shaped, The configurations may include patch-shaped, oval, star-shaped, or spherical. The device may be coiled, tubular, mesh-like, or The device may be wrapped into a nested structure. When retrieving, avoid configurations that may result in device migration from the implant site. Avoid using devices that are small enough to move through the recipient's blood vessels. Preferred embodiments of the present invention provide a high degree of structural integrity and are Such shapes include rectangular patches, disks, cylinders, and Includes flat seats.

[0119] In one embodiment, the device or assembly is retrievable after implantation, and the device Preferably, the device has a tether to aid in retrieval. Zar is well known.

[0120] In another embodiment, the device or assembly is placed at or near the desired anatomical site. Sutures placed adjacent to the patient to prevent it from migrating, moving, or crisscrossing inside the patient. Any means for suturing or securing the device or assembly may: It is within the skill of one in the art to provide such a method, for example, by attaching a suture tab to a device such as that disclosed in commonly assigned U.S. Pat. No. 29 / 423. A device or assembly similar to the device or assembly described in '365 In one embodiment, the device assembly can be fabricated in a homogenous transfer The explants are supported by a similar containment device, such as a Theracyte™ device. in unimmunized rodent recipients and human recipients It is expected to protect against rejection. Brauker et al., Neovascularization tion of synthetic membranes directed by membrane microarchitecture, J.Biomed.Mate Res., 29:1517-1524, 1995; Tibell et al., Surviva l of macroencapsulated allogeneic parath yroid tissue one year after transplantat ion in nonimmunosuppressed humans, Cell T transplant., 10:591-599, 2001; and Kumagai-Br Aescha et al., The TheraCyte™ Device Protects against Islet Allograft Rejection in Im munized Hosts, Cell Transplant., October 2012 3 Similarly, xenografts can also be implanted with the Theracyte™ device. Instead, leakage of foreign antigens can cause a strong inflammation in the vicinity of the implant. Brauker et al., Local inflammatory response sponse around diffusion chambers contain ing xenografts.Nonspecific destruction o f tissues and decreased local vasculariz ation, Transplantation, 61:1671~1677, 1996; Loudovaris et al., Destruction of xenografts bu t not allografts within cell impermeable membranes, Transplant.Proc., 24:2291~2292 ;Loudovaris et al., CD4+ T cell mediated destruct ction of xenografts within cell-impermea ble membranes in the absence of CD8+ T c cells and B cells, Transplantation, 61:1678 ~1684, 1996; and McKenzie et al., Protection of xe nografts by a combination of immunoisola tion and a single dose of anti-CD4 antib See Cell Transplant., 10:183-193, 2001. I want to be.

[0121] In other embodiments, the device assembly may include a single separator that further separates the lumen of the device. Or it may consist of two or more seals, i.e., bulkhead seals. Takumi Application Nos. 29 / 408366, 29 / 408368, and 29 / 408370 See, for example, U.S. Pat. No. 29 / 423,365. Such designs are suitable for large crowds. The densely packed cells in the center of the cluster / agglomerate are denied or lack of nutrients and oxygen to the cells. Large cell aggregates or aggregates of cells are formed, so that they have reduced access to the target cells and are therefore potentially non-viable. It does not inhibit, reduce, or promote cell clusters or cell clumps. Thus, devices containing multiple chambers or compartments allow cells to be cultured in one or more Better distribution across multiple chambers / compartments is possible. In this way, each cell has a greater opportunity to receive nutrients and oxygen, which increases the promotes cell survival rather than death.

[0122] In one embodiment, the present invention provides a device comprising a cell chamber of substantially oval to rectangular shape. These devices further relate to a device or assembly in which the weld or seam is The device runs through the center of the chair, with either a weld or a seam sealing each side of the device. This creates two separate reservoirs, lumens, chambers, and void spaces. compartmentalized or recompartmentalized to form a container or compartment Consisting of a single piece of metal, or separated or divided in the middle by a weld However, in this case, such a weld does not completely seal the chamber. The accordion-shaped chamber is made of a material with welds or seams. It is compartmentalized or reorganized to create

[0123] Another embodiment may have 2, 3, 4, 5, 6, 7, 8, 9, or From a substantially oval or rectangular cell chamber with 10 or more welds Other similar devices or assemblies (see, for example, U.S. Pat. No. 8,425,928) In some embodiments, the welds are formed on the horizontal planes or surfaces of the device. In other embodiments, the weld spans a vertical layer or face of the device. In some cases, cross welds are present across both the horizontal and vertical layers of the surface. In some embodiments, the welds are parallel and equidistant to one another. In other embodiments, the welds are perpendicular. In still other embodiments, the welds are parallel but not equidistant. As can be seen, such a design allows the welds to run across both boundaries of the device, Connect up to 2, 3, 4, 5, 6, 7, 8, 9 fully isolated , or 10 or more chambers may be effectively formed, with one continuous chamber - but interdigitated, forming separate regions within the same chamber Furthermore, it is possible to create a weld that is parallel or parallel and equidistant. Although certain exemplary devices are described, the welds may be in any direction or orientation. Still another device in accordance with the present invention is a device for forming a long weld having an area that is not interrupted by the weld. It is also possible to customize or fabricate devices that include the type of weld used and The number of cells or drugs employed and the therapeutic method or purpose for which they are employed In some embodiments, the welds are arranged to modify the appearance of the device. It can be placed.

[0124] 3 to 14 show embodiments of three-dimensional cell encapsulation devices or assemblies, As described above, these are merely exemplary embodiments, and one skilled in the art would be able to implement any such device. Use welds or seams in chairs to create different configurations These can be envisioned by the applicant, and the shape can be modified. and other features described in the preceding paragraph to form a device or assembly suitable for the intended purpose. For example, the device can be designed to detect ultrasound waves around the entire circumference. It can also be welded to create a completely closed internal lumen, allowing for multiple lumens to be The membrane can be sealed or walled to form a pouch-like device. Other means of constructing the lumen may also be used. The lumen is centrally located and runs along the length of the device. It is further compartmentalized by an internal weld that projects along the axis. , which effectively limits the thickness or depth of each compartment but does not completely isolate the inner lumen. This technique allows the width and depth of the compartment to be controlled, Can be altered as required to allow for viability and efficacy of cell products Additionally, but not limited to, overall length, overall width, perimeter weld thickness, perimeter weld Joint width, compartment length, compartment width, compartment depth, All dimensions of the device, including internal weld length, internal weld width, and port locations Methods modified to optimize devices for specific cell products and / or bioactive agents It is a design standard that can be achieved.

[0125] For example, in Figures 3-70, the compartments are assembled into the device during the perimeter ultrasonic welding. Cell products or bioactive agents are loaded through two separate ports embedded in the chamber. These ports are designed to distribute cells and / or drugs evenly during loading. For the purpose of accessing the compartment, it extends into the lumen or compartment. In certain embodiments, the port allows for loading of cells or therapeutic agents into another port. Ventilate the cell chamber while loading, thereby preventing pressure buildup within the device. This will help to stop it.

[0126] Alternatively, in another embodiment, the device or assembly presented herein comprises: It does not contain any input or output ports, i.e., the device is said to be portless. In another aspect, the perimeter welds and compartmentalized spot welds are ultrasonically welded. The spot welds act similarly to internal welds and are created by , so as to periodically limit the expansion of the lumen or compartment at any given point. Therefore, here too, the loop created by the spot welding The compartments or compartments are interconnected and no single Neither the body nor the compartments are isolated or completely separated. This can be achieved for a single cell chamber within the device, and can be done within the device or assembly. This can also be achieved for multiple cell chambers within a cell assembly, either within a device or assembly. This can be achieved for any one cell chamber within the assembly. The total number, diameter, and distribution of the dot welds will affect the loading behavior of any cell product or drug. These are design parameters that can be optimized to suit the cell growth pattern and growth rate.

[0127] Once the cells are loaded into the device, a second ultrasonic weld around the periphery of the device seals the cells. The result of the multi-step sealing process is The finished device is completely closed and no ports protrude beyond the periphery. The grooves may be peripheral areas where fractures may occur as a result of less than optimal ultrasonic welding. This approach simplifies the loading process and improves the overall integrity and Safety is improved.

[0128] Furthermore, although the above process has been described in two sequential steps, the cells and / or drugs may be The method for encapsulating the agent is not limited to the two steps described above, and may involve encapsulating the cells and simultaneously and any number of steps necessary to prevent or reduce the level of damage to the device. Steps in any order.

[0129] Those skilled in the art will appreciate that this can be done in a variety of ways, for example, as an internal shaft, where the material can be cut immediately after welding. This can be achieved by using an ultrasonic sonotrode with a tapered edge. These machined welds promote device-induced angiogenesis, which in turn reduces oxygen-dependent more readily accepted to improve survival and efficacy of cellular products and / or drugs The device has the advantage of being integrated into the patient's tissue, promoting and increasing new vasculature through the device. As a result of this, diffusion in the X and Y directions occurs, which is usually limited to the center of a planar sheet device. This improves oxygen transport through the

[0130] In other embodiments, the device design may be of a different shape, e.g., a thin The vesicle encapsulation device may be in the form of a tube or flattened tube, or the device of the present invention. It may be any other such shape that meets one of the above requirements for a suction cup.

[0131] Device Materials Useful biocompatible polymeric devices include: (a) a core containing tissue or cells; ) a biocompatible semipermeable membrane (jacket) that does not contain isolated cells (i.e., immobilizing the cells) and a peripheral or rim region of the membrane (the membrane itself, which does not undergo polymerization).

[0132] The "semipermeable" nature of the device membrane allows molecules produced by cells (metabolites, nutrients, and The device allows diffusion of the therapeutic agent (and other substances) from the device into the surrounding host tissue, but does not allow diffusion of the cells within the core. It is sufficiently impermeable to protect the cells from harmful immune attack by the recipient.

[0133] U.S. Patent Nos. 6,773,458; 6,520,997; 6,156,30 No. 5; No. 6,060,640; No. 5,964,804; No. 5,964,261 No. 5,882,354; No. 5,807,406; No. 5,800,529 ; Same No. 5,782,912; Same No. 5,741,330; Same No. 5,733,336; Same No. 5,713,888; Same No. 5,653,756; Same No. 5,593,440; Same No. 5,593,440; Same No. 5,593,440; No. 5,569,462; No. 5,549,675; No. 5,545,223; No. No. 5,453,278; No. 5,421,923; No. 5,344,454; No. 5 ,314,471;Same No.5,324,518;Same No.5,219,361;Same No.5, 100,392; and 5,011,494, all of which are incorporated herein by reference in their entirety. Cell-permeable and cell-impermeable membranes have been described in the art, including the patents listed in are.

[0134] (including but not limited to) polyacrylic acid (including acrylic copolymers), polyvinyl alcohol Polyvinyl chloride copolymer, polyurethane, polystyrene, polyamide, acetate cellulose, cellulose nitrate, polysulfone (including polyethersulfone), polyphospha Polyacrylonitrile, poly(acrylonitrile-co-vinyl chloride), PTFE and various polymers and polymers, including derivatives, copolymers, and mixtures of the foregoing. The blend can be used to manufacture the device jacket.

[0135] Biocompatible semipermeable hollow fiber membranes and methods for making them are each incorporated herein by reference. Incorporated U.S. Patent Nos. 5,284,761 and 5,158,881 (also In one embodiment, the device is The chair jacket is disclosed in U.S. Pat. No. 4,976,878, each of which is incorporated herein by reference. Polyethersulfones, as described in US Pat. Nos. 59 and 4,968,733 It is formed from polyethersulfone hollow fibers such as hollow fibers.

[0136] In one embodiment, the encapsulation device is made of an anisotropic material, such as, but not limited to, polysulfone. Polyimide (PSF), nanofiber mat, polyimide, tetrafluoroethylene / polytetrafluoroethylene Polytetrafluoroethylene (PTFE; also known as Teflon®), e PTFE (expanded polytetrafluoroethylene), polyacrylonitrile, polyethers In addition to cellulose acetate, cellulose nitrate, and polyamide, These membranes include biocompatible materials, including HPMC membranes. The types and compositions of substantially similar membranes include, to name a few: At least Gore®, Phillips Scientific® , Zeus®, Pall®, and Dewal® It is manufactured.

[0137] Device Loading One embodiment for loading a therapeutic agent containing cells into an implantable device or devices This application is incorporated herein in its entirety by reference to the accompanying application filed on February 21, 2011. Publication No. WO / 2012 / 115619 (PCT / US11 / 25628) by the applicants. , LOADING SYSTEM FOR AN ENCAPSULATION DEV It is listed on the ICE.

[0138] In another embodiment, cell loading with the above device involves thawing PECs and culturing them. From the time of cultivation until the cell aggregates are counted and loaded into the device, they are closed. It is fully automated so that it is contained within a closed, sterile environment.

[0139] In another embodiment, a syringe-like system is used to load the cell aggregates into the device. Do it.

[0140] These and other similar methods will be apparent to those skilled in the art.

[0141] cell density Cell loading density can be varied over a wide range for any device. The number of cells to be loaded depends on the expected dose or dose required by the treatment, and The number of macroencapsulation devices employed in the treatment will depend on the type of treatment.

[0142] In one embodiment, 10×10 3 ~10×10 9 cells into a device or assembly Each chamber (compartment or lumen) of the syringe is loaded with a bolus of 1000 mg of ethanol. So, our method for generating PECs is: Approximately 3-4 million cells, or approximately 367,000 cells per microliter In one embodiment of the present invention, approximately 250 μL of cell aggregate suspension is The total cell count of the EN250 device, which is a device capable of holding suspension, is approximately 9 cells. In another embodiment, each of the cell aggregate suspensions is about 100 μL. Cells are placed in a multiple cell chamber device (e.g., Figures 3-70) with a capacity to hold For example, the assembly consists of eight 100 μL cell chambers (or Approximately 3-4 million cells per member), or approximately 240-300 million cells The cell chamber can be any size. For example, in Figure 5, the cell chamber of the three-dimensional device is approximately 121 μL (200 μm Therefore, eight cell chambers with a capacity of approximately 121 μL each are used. The device or assembly with the 1.5 member is approximately 968 μL, with a total cell capacity of Approximately 36 to 44 million cells per chamber (367 cells per μL) 121 μL = 44.4 million cells per chamber) for 8 cells With the cell chamber, in total, approximately 10 cells per assembly as described herein can be accommodated. 354.9 million units.

[0143] The above represents the maximum volume capacity of a single device or individual cell chambers within a larger device. This describes the theoretical cell dose or number of cells based on the capacity, and not the actual dose. The amount of time may depend on the type of cells and / or medium in which the cells are placed for loading purposes. The actual cell dose may also vary depending on the device or cell type. The cell culture was measured as a percentage of the total number of cells loaded or seeded into the cell chamber. Is it a homogenous pure culture of therapeutic cells or a population of different cell populations? Similarly, for macrocellular encapsulation delivery, it is desirable to use as few devices as possible. or the fewest possible cell chambers in any device, if possible, within the device , preferably 10 or less, preferably 9 or less, preferably 8 or less cell chambers, No more than seven cell chambers in a device, no more than six cell chambers in a device, No more than five cell chambers in a device, no more than four cell chambers in a device, No more than three cell chambers in a device, no more than two cell chambers in a device, and It is preferred to implant no more than one cell chamber per cell. The number of members depends on the capacity of the chamber lumen.

[0144] Multi-chamber modular device In one embodiment, the cell-free zones are interconnected by folds and bends. A device or assembly containing a plurality or multiple cell chambers is provided. For example, one embodiment may include a plurality of porous cell chambers laterally connected to one another. In one such embodiment, the plurality of porous fibers comprises a plurality of bars (see FIGS. 3-70). The cellular chamber may be formed, for example, by arranging the top and bottom surfaces of the porous material substantially parallel to the longitudinal axis of the device. Formed by ultrasonic welding along a straight line, 2, 3, 4, 5, 6, 7 1, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 or more Each cell chamber has a fixed capacity, e.g. For example, 100 μL, with one or more ports and an internal matrix scaffold or With foam, periodically restricting the expansion of the lumen or compartment, if desired; In one aspect, the cell encapsulation device described herein has one or more internal welds. The device comprises at least two porous chambers or a cell line suitable for human use derived from pluripotent stem cells. When stored and implanted in a suitable dose of pancreatic islets, it can be used to treat and improve the condition of subjects with diabetes. In a preferred embodiment, each chamber has substantially the same inner diameter. If multiple chambers are available, the decision is made. The volume of cell preparation requested, the disease treatment prescribed, and the amount of cell preparation required are all within the knowledge and skill of the supplier. Any number or combination of chambers may be used depending on the treatment regimen.

[0145] In one embodiment of the present invention, the Adjacent cell chambers within a bri are of different designs, volume capacities, cross-sectional dimensions, and In one aspect, the polymer mesh is ultrasonically welded from the proximal end to the distal end. and creating a cell-free zone at each weld, resulting in multiple porous cell chambers. The top and bottom surfaces of the cell chamber are formed by ultrasonic welding or acellular welding. One or more cell chambers, except where interrupted by other features that create zones The core or center of each cell chamber is continuous throughout the chamber. This can occur if the cell chamber diameter is too large or too wide. The chamber is designed to reduce the possibility of a necrotic core of cells in the center of the device. A seal or a cell chamber was placed inside the cell chamber to create a "cell-free" zone in the center of the chamber. Such cell-free zones or welds may also be described in commonly owned U.S. Pat. No. 8,278,106, particularly Figures 2-7, and the previously mentioned device of the present applicant. These cell-free zones or welds are also described in the design application. , e.g., 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120 , 125, 130, 135, 140, 145, 150, 155, 160, 165, 170 , 175, and 180 degrees, thereby simultaneously Limit the footprint of the entire device assembly by the number of chambers, or in some cases More cell chambers can be accommodated in the device assembly while reducing or eliminating the This results in a configuration that increases cell volume through the addition of It is possible.

[0146] In a preferred embodiment of the invention, the device is protected by acellular zones and / or welds. See, for example, Figures 13-14. In one such embodiment, the plurality of porous cell chambers are formed on the upper surface of the porous material. and the bottom surface by ultrasonic welding along a line substantially parallel to the longitudinal axis of the device. At least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 13, 14, 15, or 16 or more cell chambers. Each chamber may contain one or more ports on the same or opposite side. It may have a structural matrix scaffolding and / or may contain internal welds. do.

[0147] Alternatively, the individual cell chambers within any device or assembly may be in the same compartment. Each chamber does not need to have the same configuration or design. Cell chambers that may contain elastomeric foam (including, but not limited to, U.S. Pat. No. 6,149,263), The cell chamber with internal weld partitions previously described in U.S. Pat. No. 8,278,106 cell chambers with different mesh outer layers; cell chambers with different porous membranes - a further porous membrane (e.g., an angiogenic membrane, which elutes certain factors that promote angiogenesis) Cell chambers with membranes for cell dosing, different sizes of cell chambers for customizing cell dosage, The device may have different characteristic designs, including multiple cell chambers. The chamber or multiple cell chamber assembly delivers high therapeutically effective doses to the patient. At the same time, it provides flexibility in the administration scheme and does not increase the footprint of the device. It is important for various purposes.

[0148] Device fabrication In one non-limiting embodiment, the outer mesh layer is (but is not limited to) cell impermeable. the porous layer, adhesive layer or film, and any other components required for the device. One or more cell chambers comprise a variety of components, including elements (e.g., ports). A manufacturing process for making one or more of the devices or assemblies comprising: The manufacturing method may include, but is not limited to, forming each of the layered components of the cell chamber. , pressing step, welding step, casting step, molding step a step of injection molding, a step of die molding and / or die punching Punching and / or cutting steps (e.g., laser cutting, water jet cutting) One or more of the layers may be aligned and pressed. They can also be machined, for example cut together with a laser.

[0149] In another non-limiting manufacturing process, one or more layers of the device are Create engineering drawings and / or computer images of one or more parts of the device One common commercially available software package is AutoCAD is the preferred choice, but other mechanical drawing software packages are available and may be used. These layers can be bonded together by (but not limited to) heat staking, welding (high frequency or or ultrasound), adhesives, taping, high pressure and high temperature melting, and conventional pharmaceutically acceptable They may be attached to one another by techniques common in the art, including bonding with adhesives, films, etc. In a preferred embodiment, the method is based on its rapidity, cleanliness (no solvents), and thinness. Use ultrasonic welding to create a narrow seam and for strength, secure the cell chamber. or bonding different flexible sheets of the device together.

[0150] Closing or sealing the device In one embodiment, the device assembly seals the cells within the cell chamber. At least one, preferably at least one, formed from a weld to adequately enclose the Both consist of two cell chambers. Many techniques are used to weld plastics together. In the present invention, any of them is a cell chamber device or an assembly. For example, the device herein , high frequency ultrasonic welding, gluing, and clamping, but do you use a heat gun? or hot air, which softens both the parts to be joined and the plastic filler rod. Bring jet, hot gas welding or hot air welding; hot air welding / hot gas welding; (it's Heat sealers, including but not limited to hot bar sealers and impulse sealers; hot air ( or inert gas) is simultaneously applied to the welding area and the tip of the welding rod, High speed tip welding; Injection welding, especially for joining materials over 6mm thick; Contact welding Hot plate welding; Radio frequency welding; Injection welding; Ultrasonic welding; Friction welding; Spin welding welding; laser welding; transmission laser plastic welding; and solvent welding (are Other plastic welding methods are contemplated, including but not limited to: These and other methods for welding blocks are well known and those skilled in the art will be able to Any means suitable for the need to adhere or join materials may be employed.

[0151] In another embodiment, any suitable method of sealing the cell chamber may be used. Preferred sealing methods include polymer adhesives, crimping, knotting and These sealing methods are known in the art and include the use of heat sealing. In another preferred embodiment, the method described in U.S. Pat. No. 5,738,673 , using any suitable "dry" sealing method. A substantially non-porous fitting is provided for introducing the cell-containing solution. After filling, Sealing the Device. Methods for sealing a device are known in the art. do.

[0152] In another embodiment, a method of closing a cell chamber or device comprises: wetting at least a portion of the permeable polymer membrane with a liquid; applying heat to at least a portion of the wetted thermoplastic polymer associated with the membrane; and creating closure using the same. The closure is called a "wet seal." In this "wet sealing" process, the thermoplastic The polymer melts at a lower temperature than the polymer film. The fluid is integrated into the polymer membrane and flows along the surface and into the open gaps in the membrane. Filling the polymer membrane completely through the closed area with the molten polymer. Once the thermoplastic polymer is cooled below its melting point, the device A closure is formed within the well. The closure is cell-tight and often liquid-tight. The portion of the device having a closure formed by a slit seal provides a cell-impermeable region of the device. is depicted.

[0153] In one embodiment, a method of closing a storage device comprises: wetting an ePTFE membrane with a liquid; A portion of the membrane associated with a thermoplastic polymer such as fluorinated ethylene propylene (FEP) is thermally and applying a pressure to the surface of the liquid to create closure. The polymer is formed by melting and fusing to itself and the membrane in the .

[0154] In one embodiment, a method for closing a cell chamber or device comprises the steps of: Sufficient heat is applied to the portion of the permeable membrane associated with the thermoplastic polymer to melt and flow the thermoplastic polymer. After heating, the membrane / thermoplastic polymer combination is entangled in the heated zone to form a closure. During the heating or entangling of the materials, the film / heat The combination of thermoplastic polymers is also stretched. After heating, entanglement, and stretching, the separated regions is formed, cutting the membrane in the separation region.

[0155] These and other "wet seal" sealing methods is described in detail in US Pat. No. 6,617,151.

[0156] Immobilization Device

[0157] In one embodiment, the encapsulated cells and / or bioactive The agent is maintained at the implantation site, e.g., by the expression and secretion of a therapeutic polypeptide. An implantable device is provided that allows diffusion from the implantation site. Such means for securing at the site may be the suture tabs on the device, as described above. However, other means of securing or adhering the device to the anatomical site are also envisioned. The implantation site is at or adjacent to the tissue or organ that is the focus of treatment. The drug delivery from the device is not location-dependent, and the drug biodistribution is In other embodiments where the blood vessels are dependent on the vasculature, the device may be placed in a location remote to the large blood vessels or capillary beds. The implant may be located at or adjacent to a preferred In embodiments, the biocompatible device is applied to the forearm, or the flank, or the back, or the buttocks, or limb, etc., if it remains substantially intact until its removal or explantation is required. It is implanted subcutaneously, under the skin at the site of the injury.

[0158] Extended Devices Conventional implantable devices are generally made from rigid, non-expanding biocompatible materials. In one embodiment, an expandable device or assembly is provided. Whether this is possible depends on the materials employed to fabricate the device, e.g. The specific properties of the polymer sheath may determine whether the device is expandable or non-expandable. The cells may also be designed to have expandable capabilities, e.g., to accommodate additional cells. Expandable devices are available to fit your existing device or to replace it. .

[0159] In one embodiment, the large capacity device or assembly is semi-rigid and non-expandable. However, one or more small or large cell encapsulation devices may be contained therein. The holder may be contained within a larger housing or holder or cage. The cassette holder resembles a cassette holder that can hold one or more cassettes. Alternatively, the holder may be a plurality of devices, only some of which may contain cells. The devices are loaded with or contain cells within them, while other devices are empty and will be used later. A device that can be loaded and filled with cells or drugs at the time of implantation or at any time after the initial implantation. Such an implantable housing may comprise an inert material suitable for implantation within the body, For example, titanium, titanium alloys suitable for implantation in a mammalian body, more particularly in a human body. Includes metals, plastics, and ceramics, which are gold or stainless steel alloys.

[0160] Refillable cell encapsulation device In one embodiment, provided herein is a refillable reservoir, lumen, container, A containment device with tenors or compartments, which periodically dispenses the appropriate therapeutic agent. The cells are loaded with therapeutic agents and / or cells or bioactive agents and / or cells. This relates to the containment device, which may be filled or flushed with such a filling device. For example, a syringe, or in vivo a reservoir, lumen, container or other standard means in the art for filling compartments etc. using a therapeutically effective amount of a suitable therapeutic agent and / or therapeutic cells or bioactive agents. The active agent and / or cells are delivered to an implanted reservoir, lumen, container, or compartment and subcutaneously (subdermally or subcutaneously) y) can be achieved by injection.

[0161] Encapsulated cells

[0162] In one embodiment, the cells encapsulated within the three-dimensional large capacity device assembly are endoderm cells. mesoderm, (but not limited to) PDX-1 negative foregut, PDX-1 positive foregut, and pancreatic Pancreatic endoderm (PE or PEC), pancreatic progenitor cells, Endocrine precursor cells (precursors or progenitors), immature beta These include (but are not limited to) definitive endoderm-lineage cells, including endocrine cells such as blastocysts. Generally, definitive endoderm-lineage cells also refer to definitive endoderm and their derivatives and progeny. Any cell derived from, including but not limited to, lung, liver, thymus, parathyroid gland, and and any cells of organs derived from the intestinal tract, such as the thyroid, gallbladder, and pancreas. Grapin-Botton and Melton, 2000; Kimelman and Griffin, 2000; Tremblay et al., 2000; Wells and Melt See Wells and Melton, 1999; Wells and Melton, 2000. These embryos Definitive endoderm-lineage cells and other definitive endoderm-lineage cells are described in detail by the applicant. and at least other suitable embodiments described herein are disclosed in U.S. Pat. No. 7,955,655. No. 8,585, PREPRIMITIVE STREAK AND MESENDODE RM CELLS; 7,510,876, 8,216,836, 8,6 No. 23,645 DEFINITIVE ENDODERM; No. 8,129,182; ENDOCRINE PRECURSOR CELLS,PANCREATIC HOR MONEEXPRESSING CELLS AND METHODS OF PROD UCTION; No. 8,278,106, ENCAPSULATION OF PAN CREATIC CELLS DERIVED FROM HUMAN PLURIPO TENT STEM CELLS; and U.S. application filed December 13, 2013 No. 14 / 106,330, IN VITRO DIFFERENTIATION OF PLURIPOTENT STEM CELLS TO PANCREATIC EN In doderm cells (PEC) and endocrine cells It is described in more detail.

[0163] The present invention also relates to the production of cells from animals, provided that the cells are capable of differentiation as defined herein. Differentiable cells derived from any source in the art are also contemplated. For example, differentiable cells may be derived from an embryo Alternatively, it can be extracted from any primordial germ cell layer therein, such as placental tissue or chorionic tissue. It can also be extracted from tissues, including (but not limited to) fat, bone marrow, neural tissue, and breast tissue. , adult stem cells including liver tissue, pancreas, epithelium, respiratory tissue, gonadal tissue, and muscle tissue. In a specific embodiment, the differentiable cells are , embryonic stem cells. In another specific embodiment, the differentiable cells are adult stem cells. In yet another specific embodiment, the stem cells are placenta-derived stem cells or chorion-derived stem cells. be.

[0164] Of course, the present invention applies to cells derived from any animal capable of giving rise to differentiable cells. The animals from which the differentiable cells are obtained may be vertebrates. may be invertebrates, may be mammals, may be non-mammals, may be humans Examples of animal sources include (but are not limited to) primates. Animals, including rodents, dogs, cats, horses, cattle, and pigs.

[0165] The differentiable cells can be derived using any method known to those skilled in the art. For example, human pluripotent cells can be generated using dedifferentiation and nuclear transfer methods. Therefore, as used herein, human ICM / epiblast cells or primitive ectoderm cells are in It can be derived in vivo or in vitro. As described in WO99 / 53021, primitive ectodermal cells can be cultured in adherent culture medium. They can be grown in culture or as cell aggregates in suspension culture. Furthermore, human pluripotent cells can be cultured by manual methods, such as enzymatic or non-enzymatic passaging. using any method known to those skilled in the art, including subculture by Subculture is also possible.

[0166] Embodiments of the compositions and methods described herein include, but are not limited to, Cy T49, CyT212, CyT203, CyT25 (at least at the time of filing this application) 3550 General Atpmics Court, San Diego, C. (commercially available from ViaCyte Inc., A 92121), BGO1, Human pluripotent stem cells, such as hESCs, including BG02 and MEL1, as well as iPSCs 482c7 and iPSC-603 (Cellular Dynamics Inter National, Inc., Madison, Wisconsin), and iPS iPSC-G4 (hereinafter referred to as "G4") and iPSC-B7 (hereinafter referred to as " B7”)(Shinya Yamanaka, Center for iPS Cell Induced pluripotent stem cells (i Various types of cells, including induced pluripotent stem cells (PS) The use of differentiable primate pluripotent stem cells is envisaged, and G4 and B7 are used herein. Some of these human pluripotent stem cells are being developed in the United States. It is registered by a national registry, such as the National Institutes of Health (NIH), and is registered with the NIH Human Stem Cell Registry (e.g., CyT49: registration number 0041) Information on another available cell line, CyT49, is also available in the stemcells.nih.gov / research / regist online ry. Still other cell lines, such as BG01 and BG01 v and Wisconsin International Stem Cell WiCell (registered trademark) (catalog name: BG), a subsidiary of (WISC)Bank 01) and sold to third parties by ATCC (catalog number SCRC-2002) Other cell lines described herein are available from WiCell® or cannot be deposited or distributed in a biological repository such as ATCC, Such cell lines are available directly or indirectly from leading researchers, laboratories, and / or research institutions. are available to the public indirectly, e.g., through public requests for cell lines and reagents. These cell or material transfers are common knowledge to those skilled in the art of life science. or via a standard material transfer agreement between the material owner and recipient. These types of mass transfers occur frequently, especially in life science research environments.

[0167] In August 2006, Klimanskaya et al. reported that hESCs can be derived from a single blastomere. This keeps the embryos intact and does not cause their destruction. Using micromanipulation techniques, biopsies were taken from each embryo and 19 ES cell-like cells were isolated. We obtained a single embryonic stem cell line and two stable hESC lines. These hESC lines were cultured in an undifferentiated state for 6 months. It can be maintained for more than a month, and Oct-4, SSEA-3, SSEA-4 , TRA-1-60, TRA-1-81, Nanog, and alkaline phosphatase These hESCs showed normal karyotype and expression of pluripotency markers, including pluripotency. This is possible both in vitro and in vivo in teratomas. , forming derivatives of all three embryonic germ layers. New stem cell lines without embryo destruction. These methods of creating human embryos address the ethical concerns of using human embryos. nskaya et al. (2006) Nature, 444:481-5, Epub, 2006 See August 23. However, Klimanskaya et al. The line was co-cultured with other hESCs. Later, in 2008, Chung Y. et al. hES cell lines could be derived from single blastomeres but without co-culture with hESCs . Chung Y. et al., Cell Stem Cell, 2008, 2(2), 113~ 117. Thus, the preparation of cells for encapsulation as presented herein involves: It can be done without the destruction or commercialization of human embryos.

[0168] Databases exist that describe and provide information about a variety of pluripotent stem cell lines. These databases are provided by the National Institutes of Health (NIH) and are updated regularly. IH) Human Stem Cell Registry, Human Embryo nic Stem Cell Registry and University of M Assachusetts Medical School, Worcester, Ma. International Stem, located in Sachusetts, USA Including (but not limited to) Cell Registry.

[0169] Methods for increasing cell viability

[0170] Adequate oxygen and oxygen supply throughout the polymer membranes used to encapsulate cells and tissues. Lack of nutrient transport is one obstacle to the field of cell encapsulation and tissue encapsulation / immunoisolation. The result of this inadequate gas and nutrient exchange is a decrease in metabolic activity. and cell death. The embodiments described herein address this deficiency in the prior art. This invention relates to an implantable cell encapsulation device.

[0171] The oxygen tension within the pancreatic islets varies in their native environment, after isolation, and in various potentiated islets. after implantation in a renal device, as well as in a naked or free state, e.g., under the kidney capsule. The partial pressure of oxygen in the pancreatic islets is the highest of any organ in the body (37–4 6 mmHg). However, when isolated, these values ​​are significantly reduced (14-19 mmHg). When islets are transplanted into normoglycemic animals, the values ​​are somewhat smaller compared to their isolated values. (9-15mmHg). Dionne et al., Trans. Am. Soc. Artf. Intern.Organs., 1989;35:739-741; and Carlss See on et al., Diabetes, July 1998, 47(7):1027-32 These studies have shown that tissues, when immunoisolated and transplanted, can migrate to vascularized areas such as the kidney capsule. Even within the stratum corneum, the oxygen partial pressure drops compared to its natural state (37-46 mmHg). Therefore, it is particularly useful to place cells in the core of a cell cluster or in an encapsulation device. The closer to the core, the more likely these near-anoxic conditions will result in cell death. do.

[0172] Good oxygen availability to encapsulated cells or tissues and / or bioactive agents To achieve this reliability and delivery, the embodiments described herein may be implemented in a variety of ways, e.g., The design and / or construction of the device, e.g., the membrane or This document relates to the use of perfluorinated substances in materials and in particular to perfluorinated organic compounds, e.g. For example, perfluorocarbons (PFCs) are those acids It is a good solvent for oxygen because its solubility is several times higher than that of water. Liquid PFCs contain 40-55% oxygen by volume and 100-150% CO2 by volume. PFCs are often used as blood substitutes and for tissue preservation. In addition, PFC derivatives are dense, chemically inert, and water-insoluble, and cannot be metabolized. It is a compound with

[0173] In one embodiment, enhanced O2 delivery is achieved by administering a PFC emulsion or some emulsion of a PFC. For example, the components or cells of the device may be mixed with an emulsifier. Coatings can be suspended, immersed, or incubated in a solution / matrix. Furthermore, certain PFCs with high weight concentrations per unit volume can be formed. emulsions are known to improve oxygen delivery and retention properties. The high oxygen partial pressure created by the O2 retention capacity of FCs reduces the amount of dissolved oxygen reaching the tissues. An O2 pressure gradient is created that drives diffusion, thereby enhancing O2 delivery to cells.

[0174] PFC substances include (but are not limited to) perfluorotributylamine (FC-43 ), Perfluorodecalin, Perfluorooctyl Bromide, Bis-perfluorobutyl ethane, or other suitable PFC. Preferred PFCs include about 60 to about 76 weight percent Perfluorinated fluids typically contain carbon-bonded fluorine. It can be a compound, but is usually a mixture of such compounds (U.S. Pat. No. 2,500,388, No. 8 (Simons); No. 2,519,983 (Simons); No. 2,594, No. 2,616,927 (Kauck et al.); and No. 4 ,788,339 (Moore et al.). PFCs are also listed in the Encyclopedia of Chemical Technology gy, Kirk-Othmer, 3rd edition, vol. 10, pp. 874-81, John Wil Also included are PFCs listed in Fischer & Sons (1980). For example, useful PFCs include: C is perfluoro-4-methylmorpholine, perfluorotriethylamine, perfluoro perfluoro-2-ethyltetrahydrofuran, perfluoro-2-butyltetrahydrofuran, Perfluoropentane, perfluoro-2-methylpentane, perfluorohexane, Perfluoro-4-isopropylmorpholine, Perfluorodibutyl ether, Perfluoro Preferred inert fluorocarbons include fluoroheptane, perfluorooctane, and mixtures thereof. Fluorochemical liquids include perfluorohexane, perfluoro-2-butyltetrahydrofuran, Includes furan, perfluoroheptane, perfluorooctane, and mixtures thereof. Commercially available PFCs useful in the embodiments described herein are 1990 product t bulletin #98-0211-5347-7(101.5)NPI, FLU FLUORINERT fluids, such as those listed in FLUORINERT fluids For example, FC-72, FC-75, FC-77 and FC-84 (Minnesota Minnesota ning and Manufacturing Company, St. Paul, M. inn.), as well as mixtures thereof.

[0175] Lumen matrix, foam, or scaffold

[0176] In one embodiment of the present invention, the supply and solubility of oxygen is increased through the containment chamber. strategies that increase oxygen solubility in the vicinity of the interior of the cell chamber. do.

[0177] In one embodiment, the method or means for increasing oxygen to the core of the cell chamber comprises , a lumen or chamber between the walls of the cell encapsulation device forming a cell chamber The method comprises applying a matrix, foam, or scaffold or insert of the The matrix is ​​essentially a matrix in which cells or cell clusters or cell aggregates are Interconnected cavities or spaces of a size that allows them to reside within the ostium or stoma. The foam matrix is ​​made up of porous or small holes that allow oxygen and other nutrients to reach the cells. Pore ​​size, pore density, and The void volume of the foam scaffold can vary. The pore shape can vary from single cells, cell clusters, or round, oval, or irregular shapes capable of holding cell aggregates. Because the shape of the ostium can vary greatly, its dimensions can vary along the axis along which it is measured. For purposes of the present invention, the pore diameter of at least some of the pores in the foam is between 40 and 100 less than 0 μm, preferably 50 to 500 μm, preferably 50 to 400 μm, preferably 5 0 to 300 μm, preferably 50 to 200 μm, and more preferably 50 to 100 μm. In one embodiment, the foam pores are circular and / or non-circular, If circular (e.g., oval), the pore dimensions are determined by the size of the pores, which determines whether the cells are in the cavity or in the pore. It can be variable as long as it is sufficient to allow retention on the surface within the pores. In addition to the cell-accepting pore size described above, it is preferable to have at least some voids within the foam. and pores that are cell-non-permissive but still allow nutrients, including oxygen, and It should be less than 10 μm in diameter to provide channels for transporting bioactive molecules. do.

[0178] The pore density of the foam (i.e., the number of pores that can accommodate cells, as described above) The number per unit volume can vary from 20 to 90%, preferably from 50 to 70%.

[0179] In one embodiment, the lumen matrix or foam is an elastomeric matrix. A variety of elastomeric polymers have been described, for example, Stabler WO2010 / 121024 by et al. discloses a complex for delivering oxygen, (including but not limited to) silicone, polyolefin, polyester, polystyrene, and a biocompatible polymer support comprising a copolymer of the formula (I) and a mixture thereof. The polymer support further includes a polymer precursor including a monomer, an oligomer, and the like. Precursors formed from, but not limited to, vinylsiloxysilanes, alkylsiloxysilanes, Siloxysilane-containing polymers, including silanes or alkylarylsiloxysilanes, and (but not limited to) polydimethylsiloxane (PDMS), polydimethylsiloxane polydimethylsiloxane monoacrylate, polydimethylsiloxane monomethacrylate, and Polymers containing mixtures may also be included.

[0180] As used herein, "silicone elastomer" or "silicone composition" or The term "silicone matrix" is a broad term and is understood by those skilled in the art. It is given its usual and customary meaning (not limited to a particular, specialized meaning) and is not limited a composition comprising a polymer having at least silicon atoms and oxygen atoms in the skeleton without Point to something.

[0181] In another embodiment, the other non-bioabsorbable material is polyethylene, polyvinyl acetate, polymer Methyl acrylate, silicone, polyethylene oxide, polyethylene glycol, polyurethane Tan, polyvinyl alcohol, natural biopolymers (e.g., cellulose particles, chitin, keratin, silk, and collagen particles), and fluorinated polymers and copolymers - (e.g., polyvinylidene fluoride, polytetrafluoroethylene, and hexafluoroethylene) The polymer may include a polymer such as propylene.

[0182] In another embodiment, PDMS is coated with an oxygenated PFC as described above as a source of oxygen. Or it can be formulated with calcium hydroxide.

[0183] Foam scaffold synthesis in general

[0184] The foam scaffold is adapted to fit the device as needed. In the (or "hollow fiber") embodiment, the foam scaffold is formed with a hollow fiber lumen. It may form a cylindrical tube or rod, as long as it can fit, and a rectangular It may form a tube or rod of any other angled shape. In some embodiments, the foam scaffold has a burr (f) that may contact the inner wall of the hollow fiber. It is envisioned that the flexure may have protrusions (ins) or other protrusions.

[0185] In one embodiment of the present invention, the cell device is constructed from a hollow fiber membrane with a cylindrical inner foam scaffold. It is formed from.

[0186] The device may also be in the form of a flat sheet device. The patents are incorporated herein by reference in their entirety. Nos. 368, 29 / 408370 and 29 / 423365, as well as already The invention is described in detail in the Baxter publication cited above. The flat sheet device generally comprises a first flat sheet membrane with a first interior surface and a second and a second flat sheet membrane with an inner surface of the membrane, the two membranes being sealed at the inner periphery. The membrane is then inserted into the ring and a foam scaffold is placed between the membrane. Cells can be introduced through it and a plug can be inserted into the port to complete the seal.

[0187] The devices of the present invention can be formed according to any suitable method. , a foam scaffold may be pre-formed and a pre-fabricated jacket, e.g., a separate structure, may be used. It can be inserted into a hollow fiber membrane as a component.

[0188] In vivo imaging capabilities

[0189] In one embodiment, cells within the encapsulation device can be imaged or visualized in vivo. Imaging plays an important role in stem cell therapy. For example, non-invasive forms of imaging can be used to (1) visualize the cells being treated and / or (2) determine the presence, severity, or phenotype of disease; (3) determine the severity, severity, or phenotype of disease; and (4) determine whether engraftment cell therapy is associated with harmful cell types or Monitor for the appearance of non-target cell types and structures such as cysts or microcysts; ) guide the delivery of treatment; and (4) track the time course of disease and assess the effect or effectiveness of treatment. (5) to name and define the therapeutic mechanism; (6) to analyze and characterize the survival and function of engrafted cells. (7) detect and monitor device-induced angiogenesis, which is critical for the survival of encapsulated cells; (8) for example, engraftment, survival, and replacement of pancreatic progenitor cells and / or Cellular therapies, including those described herein, for treating diabetes by implantation The method can be used to generally facilitate any cell therapy process by determining the local function of In addition, cell therapy aims to reduce morbidity / mortality. However, the non-invasive imaging methods described herein and in more detail below can be used to, for example, For example, it may also be used as a useful surrogate endpoint in pilot studies or preclinical studies. It can be done.

[0190] Any in vivo imaging technique must be i) non-invasive; ii) reliable; iii) be capable of tissue penetration to a depth of at least 3 mm; iv) the resolution is 500 μm or less, preferably 50 to 100 μm or less; Imaging is not attenuated by the device material, e.g., imaging through PTFE vi) clinically compatible and not technically cumbersome or complex; i) commercially available; viii) FDA approved for human use; x) Reasonably cost-effective; x) Reasonably long (e.g., seconds or minutes); Ideally, imaging of cells within a cell is possible, or any combination of the above. It is target.

[0191] Currently, state-of-the-art methods include (but are not limited to) confocal microscopy, two-photon microscopy, Microscopy, high-frequency and low-frequency ultrasound, optical coherence tomography (OCT) coherence tomography, photoacoustic tomography (PAT) oustic tomography, computed tomography (CT) tomography, magnetic resonance imaging (MRI), single-photon emission computing These include single-energy imaging (SPECT), positron emission tomography (PET), and positron emission tomography (PET). Alone or in combination, these provide a useful means for monitoring transplanted cells. Furthermore, as these technologies improve over time, it is difficult to know how each technology works. It is also expected that the basic principles of how the technology works or its usefulness will be substantially similar. In this sense, the in vivo imaging described herein is It is not intended to be limited to techniques subsequently discovered and described herein. It also includes techniques described as providing the same utility.

[0192] In one embodiment, the imaging technique employed is non-invasive and provides three-dimensional tomographic data. It provides high temporal and spatial resolution, enables molecular imaging, and is inexpensive and portable. Currently, no single modality is ideal (discussed in more detail below). Each modality has different attributes, and these modalities are complementary. This can provide complimentary information.

[0193] Confocal microscopy is a technique that increases the contrast of micrographs and uses a spatial pinhole. It is used to reconstruct a 3D image by eliminating out-of-focus light in specimens thicker than the focal plane. It is an optical imaging method that allows the creation of images with only one point in the sample illuminated at a time. Therefore, 2D or 3D imaging does not provide a uniform raster image within the specimen. (i.e., a rectangular pattern of parallel scan lines). Three main scanning variations are employed to produce confocal microscopy images. Point actuation is achieved by a fixed illumination light beam (scanned by the stage) coupled with a lateral displacement. A scanning optical beam (scanning by beam) with a moving specimen stage and a fixed specimen stage is adopted. This can also be achieved by using a spinning Nipkow disk or Nipkov The specimen is scanned by a series of light spots transmitted through an opening in the disk, and the stage and This can also be achieved by keeping both light sources fixed. Each technique has its own special It has performance features that make it advantageous for confocal applications, but this does not limit its usefulness for other applications. Limited usefulness.

[0194] All confocal microscopes use optical sectioning and imaging in sequence through relatively thick sections or whole-mount specimens. The technique relies on the ability of the technique to produce high-resolution images that are called optics. Based on cross-sections, fixed and stained specimens can be imaged in single wavelength illumination mode, dual wavelength illumination mode, Data can be collected in dual, triple, or multi-wavelength illumination modes, Images collected with different illumination and labeling strategies are consistent with each other. Live Cell Images It allows for time-lapse sequences and digital image processing techniques to be used for image sequencing. By applying this to the field of view, it is possible to display the specimen in three dimensions along the z-axis, as well as to display 3D data. It is also possible to display the time series as four-dimensional imaging. The above confocal microscopes are also intended to encompass other confocal microscopes discovered today or in the future. The use of a spot microscope is not limiting.

[0195] For example, Celltracker, Dil, and nuclear vital dyes are relatively simple protocols. Once incorporated into the molecule, certain cell surface markers and / or proteins, as well as A large number of fluorescent probes are available that can stain intracellular organelles and structures. Fluorescent markers that bind specifically, directly or indirectly, to cell surface markers of a species are This may be useful, among other things, for example, in identifying unwanted cell types. , real-time in vivo imaging for the presence of encapsulated pluripotent cells hES or human embryonic gonocytes or induced pluripotent stem cells (IPS) or parthenogenetic cells These have provided a means for detecting teratoma formation caused by pluripotent stem cells, This offers the potential for preventing these. Escaped or leaked (or unencapsulated) pluripotent stem cells can also be identified. Identification of such cells also involves the use of fluorescently labeled pluripotent stem cells whose expression is upregulated in pluripotent stem cells. This can also be done using the promoter genes OCT4 and NANOG. Similarly, certain intracellular proteins label the nucleus, Golgi apparatus, endoplasmic reticulum, and mitochondria. Fluorescent markers, as well as fluorescently labeled phalloidin that targets polymerized actin within cells These dyes are also commercially available and provide crucial information about cell fate. Ugh.

[0196] In another embodiment, two-photon excited fluorescence (TPEF) Fluorescence microscopy is a non-invasive means of monitoring differentiation. Conversely, the product cells that are not differentiated and are encapsulated within the devices described herein A very small percentage of these cells are inadvertently implanted with pluripotent stem cells (e.g., hE It is a non-invasive method to identify SC or IPS cells or parthenogenetic cells. Fluorescence microscopy relies substantially on endogenous sources of contrast, but also on, e.g., Fibrous matrix molecules can also be detected via second harmonic generation. The mirror method relies on fluorescence emission similar to that employed by confocal microscopy. (2007) used TPEF to study the biological properties of differentiating and non-differentiating stem cells. We note that quantitative differences in chemical state and shape can be revealed in two dimensions (2D). See Rice et al. (2000), the disclosure of which is expressly incorporated herein by reference. 7), J Biomed Opt., November-December 2007, 12(6) In one embodiment, the pluripotent stem cells are transfected with a fluorescent protein, such as enhanced green fluorescent protein. The vector can be genetically engineered to express a pluripotent stem cell promoter (e.g., OCT4 or NANOG, or any other pluripotent stem cell promoter identified below. It can be driven by a motor. It can be placed deeper than a subcutaneous implant, i.e., on the surface of the skin. For these deeper implanted devices, two-photon imaging is a non-invasive technique, known as confocal microscopy. Furthermore, the photon energy required for fluorescence excitation is occurs only in the focal plane and is not sensed by cells or tissues in the extrafocal plane, When used, it is less harmful to living cells than exposure to visible or ultraviolet light.

[0197] In yet another embodiment, ultrasound is portable, essentially harmless, and versatile. and upon implantation of the encapsulated cell product and / or the encapsulated bioactive agent, It can also be run in real time as a monitoring tool over the course of the implant. In particular, conventional low frequency ultrasound and / or corresponding high frequency ultrasound can be used. can be used to provide qualitative as well as quantitative spectroscopic data. High-frequency ultrasound (20-50 MHz, 30-80 μm) is used for clinical low-frequency ultrasound ( Increased imaging resolution compared to 80 μm to 1.5 mm at 1 to 20 MHz However, its use is limited to superficial tissues due to limitations in tissue penetration depth. High-resolution imaging is currently limited to anatomical locations in longitudinal mammalian studies. It allows for in vivo assessment of structure and hemodynamic function. For example, VisualSo Vevo by nics is a clinical trial that provides: (1) information on disease progression and regression in individual subjects; (2) the ability to conduct longitudinal studies; (3) the ability to study anatomical and physiological structures down to 30 microns; (3) image-guided visualization of needle injection and extraction; (4) Microcirculatory and cardiovascular blood flow assessment; (5) User-oriented devices and (6) high throughput via a research-driven interface; and (7) comprehensive measurement and It provides an open architecture that allows for data mining and annotation as well as offline data analysis. The ability to assess blood flow in the microcirculation and cardiovascular system is crucial for determining cell viability, e.g. This helps determine O2 flow and delivery. In contrast, low frequency ultrasound (approximately 7-1 0 MHz) correlates with histological cell death in acute myeloid leukemia cells exposed to chemotherapy It has been shown to detect microstructural changes in tissues associated with vascular disease. ional low-frequency ultrasound detection in apoptosis, Proceedings of the America n Institute of Ultrasound in Medicine, Ne w York, NY, 2007 (AIUM Laura MD, 2007), S18 Please refer to page 5.

[0198] In another embodiment, a contrast agent is used to differentiate between healthy and diseased tissue by magnetic resonance imaging. In yet another embodiment, magnetic resonance imaging (MRI) can be utilized. Computerized tomography (CT) or CT scan Using this technology, detailed images of tissues and structures within the body can be created. Again, contrast agents are utilized, and their specific absorption rates facilitate visualization of abnormal tissue. One use of contrast agents such as indium-111 (I-111) oxine has a half-life of In yet another embodiment, the use of positive Positron emission tomography (PET) scanning can be used to detect positron-emitting molecules, e.g., to name a few. Emissions from carbon, nitrogen, and oxygen can be measured, yielding valuable functional information In yet another embodiment, optical coherence tomography (OCT) or photoacoustic tomography (PAT) may also be used. It can also be used to study cells and tissues inside and outside the device. While CT detects differences in the reflectivity of various tissues, PAT uses low-energy lasers. The ultrasound wave produced when tissue is heated by exposure to the laser light is detected.

[0199] Employing a variety of methods and techniques or tools, alone or in combination, Implanted cells inside the device can be visualized, analyzed, and evaluated in vivo These and other techniques now known or later developed may be used in conjunction with These techniques are intended to allow for the in vivo administration of the cells and / or agents described herein. It should be utilized to the extent that imaging and in vivo monitoring are possible. can. [Example]

[0200] Example 1 Extrapolation of the PEC of the therapeutically effective dose To help ensure adoption by the patient population, the encapsulated cells contemplated by the applicant Encapsulated cell therapy for treating diabetes, such as a cellular therapy, preferably involves treating a patient with the disease. The minimum number of macroencapsulated cell products (also referred to as "VC combinations") required to deliver a therapeutically effective dose of The product must consist of a mixture of the above-mentioned components (also called "products").

[0201] In patients with insulin-dependent diabetes, the rate is approximately 200,0 00 "IEQ" (or islet equivalent) is required. IEQ is The number and diameter of islets present in the preparation, mathematically corrected for islet volume. Calculations are based on the number and diameter of the islets. Islets are approximately 150 μm in diameter. islets with a diameter of 150 μm were mathematically corrected for their volume to obtain an IEQ Normalize against the number of therapeutic IEQs treating patients with insulin-dependent diabetes mellitus. The number of cases is estimated to be 100% for islet allografts (e.g., cadaveric donor islets), islet autografts, and diabetes. The beta cell mass at the time of onset of the disease has been determined based on available information. In transplants, the target IEQ is approximately 10,000 IEQ per kilogram of body weight, but in islets Only about 40% of them survive, so about 4,000 IEQ per kg. In autologous islet transplantation, whole islets isolated from the patient's pancreas are used. The total islet volume delivered to the dorsal region is generally between 200,000 and 300,000 IE. Q (total amount delivered), which often makes patients independent of exogenous insulin Similarly, islet survival in autologous transplantation is also an issue. 05), Current Status of Clinical Islet Tra Transplantation, Transplantation, 79:1289~129 Please refer to 3.

[0202] In view of the above, the applicant believes that the total therapeutic IEQ required from the PEC graft is approximately 2 Figure 1 depicts beta cell mass on the left and On the left, graphs depicting IEQs equivalent to those of patients with similar beta cell mass and / or function are shown. The onset of diabetes occurs when beta cell mass is approximately 10-20%. However, to achieve insulin independence or disease remission, 100% of beta cells A wide range of therapeutic options are available to avoid the need to restore normal blood glucose levels (the "normal," non-diabetic state). The implanted PEC cells do not contain pancreatic islets and therefore Since the IEQ is not equivalent to the IEQ until after maturation in vivo, Applicant Encapsulated PEC grafts (or VC-01 grafts) explanted after in vivo maturation beta cell proliferation achieved in vivo based on total C-peptide protein content C-peptide was also measured in known IEQ numbers to assess the C-peptide content of the graft. The amount is extrapolated to IEQ. Figure 2A shows the total human C-peptides from various numbers of human cadaveric islets. 1 is a graph showing the protein content of the defined IE in the range of 500 to 5000 IEQ. Q aliquots of human islets were obtained from a third-party source. Total human C-peptide content was , measured using ELISA and shown in Figure 1A, human islet count (IEQ) and total CpE. A linear relationship is observed between the concentration of ATP and the peptide protein content (pM).

[0203] Using this defined linear relationship between IEQ number and C-peptide protein content, Applicant has demonstrated that the total C peaks remain constant within the range of approximately 1400-2000 pM over a period of 4-11 months. The total human C-peptide protein content of the encapsulated PEC grafts was measured, resulting in the C-peptide content level. See Figure 1B. These total human C-peptide content levels were then analyzed. Figure 1A shows the range of IEQ delivered by the VC-01 product at maturity. Specifically, the mature VC-01 translocation rate in these 60 animals can be determined. Figure 2B confirms the 25th percentile and median values ​​of total human C-peptide found in explants. See dashed line. Correlating with Figure 1A, these C-peptide levels were significantly higher in the VC-0 This corresponds to approximately 2500 to 3500 IEQ delivered by one implant. The enclosed device (having a functional volume of approximately 20 μL, or EN20 device) is In this study, the beta cell mass was approximately 2,500-3,500 IEQ, or 80 per kg. The functional capacity of the encapsulated device and the proportionately greater therapeutic capacity may be achieved. Assuming a linear relationship between the capacity to hold the volume of cells used, large drug delivery devices For example, a device holding a functional volume of approximately 250 μL (EN250 device) Approximately 12.5 times that of the EN20 device, with a maximum of approximately 30,000-45,000 IEQ. Similarly, the EN100 device (6.5 times that of the EN20 device) can have a maximum of approximately 16 It can contain 250 to 22,750 IEQ and contains four cell chambers. EN-LC (large capacity) device (EN20 device) (48.4 times the amount of the chair) may contain a maximum of approximately 121,000 to 169,400 IEQ. Thus, to deliver a therapeutically effective dose to a patient, at least about 4, about 5, Uses approximately six, seven, or eight EN250 devices or two EN-LC devices It is anticipated that a suitable encapsulation will be required to deliver a sufficient amount of PEC. The service is described in more detail below.

[0204] Example 2 3D high-capacity device assembly to optimize surface area and cell capacity for cell therapy Lee In view of Example 1, applicants have developed a large cell density that increases the functional cell dose per device. While setting out to establish a capacity device, it is also important to ensure that the device has a minimum effective area ( or footprint), e.g., occupying the smallest amount of space possible on an anatomical site within the human body. It was restricted to

[0205] The applicant's patented drug delivery device is incorporated herein by reference in its entirety. U.S. Design Patent Application Nos. 29 / 408,366; 29 / 408,368; and and No. 29 / 408,370; and No. 29 / 408,370 filed May 31, 2012. No. 423,365; and U.S. Patent No. 8,278 issued October 10, 2012. , No. 106, ENCAPSULATION OF PANCREATIC CELLS DERIVED FROM HUMAN PLURIPOTENT STEM CELL It has already been described in S.

[0206] The effective or footprint of a device is its two-dimensional area defined by the x and y dimensions. and the two-dimensional surface occupied by the device, e.g., the surface occupied by the device within the human body. The devices described above are two-dimensional, flat, or planar. It is planar and therefore has certain size constraints when considering its use in humans. For example, such devices can accommodate larger cell product or cell volumes. To achieve this, the size of the pancreas has been increasing (increasing effective area or occupied area). It is well known that islets have extremely low proliferative capacity and undergo extensive cell loss upon transplantation. Korsgren et al. (2005) supra reported that the survival rate of transplanted human islets is approximately 1 Only 0% to 20% of the patients were transplanted because the islets were in direct contact with ABO-compatible blood. Within minutes, leukocytes appeared to infiltrate the islets, indicating an immediate blood-mediated inflammatory response (IBMI). R) and result in cell loss, resulting in a thrombotic / inflammatory response. The same islet cell loss has also been reported in experimental studies. It has also been reported after transplantation of rodent islets and human islets. See N. et al., supra, p. 1291. Early work by the present applicant has The PEC cells had the potential to proliferate and the number of cells initially seeded in the device ( (e.g., 1 million, 1.5 million, 3 million, 4.5 million, 6 million, or 9 million cells) Regardless, they were shown to proliferate and mature into insulin-secreting cells. See, e.g., U.S. Patent No. 8,278,106. Thus, the presence of The size, design, and construction of the device limit and determine the number (or dose) of cells that can be The primary constraint on maximum cell number is the number of cells loaded onto the device. is not a question of cellular capacity, but of physical device capacity.

[0207] 3-70 illustrate various embodiments of the large capacity device assembly. As shown, the assemblies are preferably at least two, preferably three, per assembly. or 4, preferably 5, preferably 6, preferably 7, preferably 8, or or more cell chambers 100, or any number of multiple chambers required for a therapeutic dose. The large capacity device is three-dimensional and was released on December 12, 2011. Nos. 29 / 408,366, 29 / 408,368, and 29 / 408,369. No. 29 / 408370; No. 29 / 423,365 filed May 31, 2012 No. 8,278,106, the EN250 device or EN It is not flat or planar as already described for the 100 device.

[0208] 3-8 show an assembly of eight cell chambers 100 or compartments. Figures 3, 4, 5A, and 6 illustrate the compartmentalized lumen or The cells are formed at an angle of approximately 0 degrees between each cell chamber 100 (compared to parallel facing chambers). The fold or bend 40 is shown as an example of a folded device assembly. The bulkhead of the assembly (or the space between the compartmentalized cell chambers) Figures 7 and 8 show the cell chamber and the cell-free area. 100 are spaced apart from each other at an angle of about 20 degrees or 40 degrees, respectively. Again, the assembly is performed by bending the bulkhead or cell-free region 40. It can be bent or folded to form a three-dimensional device. The z dimension) and width (x dimension) are measured at the fold or bend 40 in the bulkhead area. For example, at 0 degrees of flexion, the overall height and width are When bent at 20 degrees, the overall height and width are approximately 10.5 mm and 25.2 mm. are approximately 9.2 mm and 44.0 mm. Therefore, the overall device assembly The effective area or footprint can be varied by changing the nature of the folds within the device. Figures 4A, 13 and 14 show the folding of the assembly. 1 shows the previous device assembly.

[0209] Generally, the cell volume or cell density inside the cell chamber in any given device is maximized. To optimize the design, the ratio of capacitance to effective area must be maximized. is the two-dimensional area, measured in x and y dimensions, occupied by a device, e.g. For example, the two-dimensional area occupied by the device inside the human body. Three-dimensional large capacity (3D EN-LC) device assembly with eight cell chambers Each cell chamber in this prototype has a volume of approximately 120 μL, and 3D EN-LC device with a maximum volume (MV) of approximately 968 μL The effective area (EA) of this 3D EN-LC assembly is shown. rea; x and y planes) is approximately 3420 mm 2 (38mm x 90mm). 3D The capacitance-to-effective area (MV / EA) ratio of the EN-LC device is approximately 0.283 (i.e., 9 68 / 3420). This is an embodiment of a planar device with a maximum capacity of approximately 249 μL, and the effective area is 2295 mm 2 (27mm x 85mm), EN250 Compare this with the capacitance-to-effective area (MV / EA) ratio of this planar EN250 device. is approximately 0.108 (i.e., 249 / 2295). The device can hold four times the capacity of a planar EN250 device, but Therefore, it does not occupy four times the effective area of ​​the 3D EN-LC device. The effective area ratio is larger (or better) than the planar EN250 device. This allows for the encapsulation of more cells over the same effective or footprint area. The 3D EN-LC device can be folded without constraints in the z dimension (height). This increased capacitance to effective area ratio can be achieved because the height is EN250 device The EN250 device has a maximum lumen diameter of approximately 1 mm. This limits the z dimension.

[0210] [Table 1]

[0211] Table 1 compares the device assemblies illustrated in Figures 3, 4, and 5A, but Without departing from the description and design described herein, the total capacitance vs. effective To increase the area ratio, angles and / or bends 40 may be created within the device assembly. Other designs employing the principles of extraction can also be achieved. For example, Figures 6-14 show Mansard (Fig. 9, 15-28), tube or series of tubes, or flat tubules (Fig. 10, 29-42), comb-like or fin-like (Fig. 11, 43- 56), wave or U-shaped or shutter (Fig. 3, 4, 5A, 6, 7), shutter (Fig. 12, 57-70), radiators, surface texturing, wafers, coils, etc. These devices are exemplified by device assemblies, which may include multiple cell chucks. Chamber 100, ports 20, 30, and various degrees of folds or bends 40 Such a design would also increase the capacitance to effective area ratio of the device. , all of these devices are embodied and described herein.

[0212] Table 2 shows the capacity versus effective area of ​​further embodiments of three-dimensional large capacity device assemblies. For purposes of comparison of different embodiments, the effective area (or footprint) is 0×20mm or 1000mm 2For the calculations in Table 2, we also keep the overall The height (z dimension) of the cell chamber is set to a lumen thickness of approximately 0.2 mm (the thickness is the same as that of the cell chamber itself). For all embodiments except flat planar devices (limited by the thickness of the device body), In addition, the 3D large capacity device assembly also keeps each cell channel constant. There is also a gap of about 0.6 mm between the members. The capacitance of each embodiment is The Roman shade design (Figure 9) offers the greatest potential for capacity. In contrast, flat (planar; 2D) devices have the potential to have the smallest capacitance. Also, the embodiment with the highest capacity-to-effective area ratio is a Roman shade device. The embodiment with the lowest maximum capacitance to effective area ratio is also a flat device.

[0213] [Table 2]

[0214] Thus, like Table 1, Table 2 also allows devices to utilize the z dimension (height). It is evident that the maximum capacitance to effective area ratio is limited to the extent that this is not possible. With this additional z-dimension and the various design configurations described herein By adding a design configuration that takes this z dimension into account, Thus, the three-dimensional large capacity device assembly herein allows for cell delivery at high cell doses or for cell therapy or cell replacement when large cell numbers are required (e.g., type 1 diabetes) It can be optimized.

[0215] The device assembly embodiments herein generally refer to layers or components, e.g. For example, but not limited to, mesh, adhesive film, and cell-impermeable layer. Manufactured by standard cutting modalities, including laser cutting and / or die cutting. The component elements allow each of the lumens to be aligned as each component or layer is placed into the welding machine. The plates are aligned so that they fit together or can be stacked properly on top of each other. The first cell chamber is fabricated by welding all layers together to form a flat sheet. In this example, the first seal portion 10 is formed along the inner periphery of the glass plate. As part of the layer, alignment features are added that are bonded to them, and then the first Trimming at the time of welding or forming the seal, or afterwards This can promote welding accuracy. The seal is also made by high frequency ultrasonic welding. This can also be achieved using heat sealing, adhesive bonding and fastening. To form a three-dimensional assembly after creating the cell chamber, the assembly is Pre-treatment with sufficient heat allows the cell-free areas between the cell chambers to form a 3D structure. The device assembly is then The assembly is placed in a mold and heat is used to impart a substantially permanent structure. The formation of the dimensional assembly can also be accomplished in a variety of ways known to those skilled in the art. For example, the device assembly may be molded or formed (e.g., clamped). It can be injection molded or can be attached to an outer frame / material with a folded shape. The outer frame / material can be bonded in various ways (e.g., high frequency ultrasonic welding). , heat sealing, adhesive bonding, and fastening. 0 indicates that the angle of the bend or fold 40 in such construction is equal to the angle shown in FIGS. Alternatively, for example, a single large capacity device may be used. Other three-dimensional high-capacity devices have also been formed by molding chairs.

[0216] For modular fabrication, FIG. 14 illustrates the construction of any number of cell chambers 100 in a modular fashion. With any number of seals 10, such as two, three, four, five, six, seven, or eight or more. This illustrates that a desired number of cell chambers can be formed. The entire multi-chamber device is welded to form all cell chambers simultaneously. Preferably, the device assembly is a cell-cell assembly. The number of chambers in each device is selected based on the cell dose for any given patient. By building the device assembly separately, or by building each individual element within the device assembly, Alternatively, the cell chambers may be constructed separately. In fabrication, such methods do not compromise the integrity or functionality of the device assembly. High frequency ultrasonic welding, heat sealing, adhesive bonding and fastening are not permitted unless Any method available in the art can be employed, including determination of the

[0217] Additionally, acellular regions or folds within the three-dimensional construct of the device can be perforated to allow for receptor allowing the entry of host cells, e.g., blood vessels perforating from one side or sides of the device; allowing the device and cells therein to traverse through the can also be increased.

[0218] 3-70 also illustrate the device assembly similar to other planar drug delivery systems. , each cell chamber 100 of the large capacity device assembly is connected to ports 20, 30, and may contain a loading tube at one end or at each end Further examples of cell chambers include scaffolds, preferably foam or mesh scaffolds, Preferably, as discussed in more detail below and shown in FIG. 71, each cell chamber or provides increased oxygen permeation or perfusion within the core of the cell chamber, The cells may contain any internal matrix that facilitates cell survival and / or distribution of cellular products. do.

[0219] Example 3 Methods for optimizing oxygen delivery and increasing insulin production The applicant has previously demonstrated that progenitor cells of PECs are more potent than mature adult islets during and after transplantation. These findings support the idea that the cells are tolerant to hypoxic conditions, such as those occurring in the In allografts and autografts, there is a lack of angiogenesis at the time of and during transplantation and the proliferation of cells. Hypoxia is a major cell survival issue. Furthermore, improving adequate nutrients and chamber The supply of sufficient nutrients to the cells in the core of the nucleus can also be optimized.

[0220] Various matrices are being explored to improve vascularization at the recipient-device interface. However, matrices or foams inside the cell chamber are not well described. Such a matrix consists of interconnected cavities and pockets. Suitable for containing and evenly distributing cell aggregates through a cell chamber or lumen. It provides architecture while simultaneously supplying oxygen and other nutrients to cells. , and also act as conduits or channels that promote survival. At least one advantage of using as a matrix material is that the material is highly resistant to oxygen or oxygenated an oxygen conduit for transporting the suspension or oxygenated fluid to the core of the cell chamber; This is due to its high oxygen solubility, which allows it to function as a tool.

[0221] Silicone elastomer potentially improves cell survival in the core of the cell chamber To determine whether this is possible, we first investigated two silicone-derived matrices: Foams made from silicone fibers or silicone hollow fibers and silicone-based mixtures Silicone is dense (non-porous) and allows liquid to flow through the membrane. This allows for its widespread use in liquids regardless of surface tension. Silicone hollow fibers have exceptional gas transport properties (PermSe by MedArray). The PermSelect membrane module has, for example, 10 cm 2 (PDMSXA-10), 2,500 cm 2 (PDMSXA-2500), 1m 2 (PDMSXA-1.0), and 2.1m 2 (PDMSXA-2.1) various film surfaces Although this results in a dense bundle of uniformly spaced hollow fibers with a high membrane surface area, other nominal membrane surface areas can also be produced. These silicone hollow The threads were assembled into a mat for attachment to the device. Table 4 below shows the mat 71A-B show the dimensions of the components of the hollow fiber, and how the hollow fiber is made of polyester yarn. These images show how the fabric is woven or knitted together. The silicone fiber mat can be made as a single or multiple layer, and a single mat The mats can then be cut into pieces to form devices, e.g., EN20 devices. The device was then fitted inside the lumen of the vise. As previously described by the applicant, the device contains approximately 6 million PECs. and implanted into mice.

[0222] [Table 3]

[0223]

[0231] Similar to the silicone hollow fiber mat, a silicone-based The foams were also prepared. The method is described in U.S. Patent No. 7,192,450 by Dexcom, Inc., SM TEC 5,624,674 and 5,605,693 by PHNOLOGIES Those skilled in the art will appreciate the techniques employed in creating the structure of the preferred embodiment. Still other methods are known for making other types of biostable foams that may be used. For example: U.S. Patent No. 3,929,971 to Roby describes a calcium carbonate coral material. , converting it to hydroxyapatite while simultaneously preserving the unique microstructure of the coral material. The present invention discloses a method for producing a synthetic membrane having a porous microstructure, which is produced by Another example is U.S. Patent No. 6,520,997 to Pekkarinen. discloses a photolithography process for creating porous membranes. In one embodiment, about 500 grams of sugar crystals are mixed with about 15 grams of water for about 3-6 minutes. The sugar crystal size (e.g., about 90 to about 250) was adjusted to form a foam. The average diameter of the sugar in microns and the amount of water added to the sugar before casting into the mold were varied. By doing so, different architectures or cavity sizes can be obtained. The mixture was pressed into a 6-well tissue culture dish to serve as a mold and incubated at room temperature overnight. For example, by baking at a suitable temperature for a suitable time, sugar crystals were dried. Other methods of drying the mixture may also be employed. and a two-component platinum-cure silicone elastomer (NuSil S) was applied according to the manufacturer's instructions. Ilicone Technology: Part Number MED-6015) at a ratio of approximately 10:1 Mix components A:B in the ratio and apply to the surface of the sugar mold at a rate of approximately 3 grams per 962 mm2 of the surface area of ​​the sugar mold. The silicone was applied evenly to the mold at a ratio of 1:1. Vacuum was applied to the mold for approximately 6 minutes. Or, for a suitable period of time, allow all the silicone to fill the cavity in the mold. The silicone is drawn into the mold through a tube and heated at about 40°C, or at any suitable temperature. The sugar template was then dissolved using demineralized water, resulting in a 6-well plate. A shallow cylindrical porous silicone (foam) well in the shape of a tissue culture well was created. 71C is a three-dimensional matrix of silicone inserted between membranes that form cell chambers. FIG. 71C shows a cross section of one embodiment of a lumen matrix comprising a foam or a tubing. The foam may also have a plurality of interconnected cavities or open spaces, the cavities being generally The layers may be formed in layers having different cavity sizes. Also shown.

[0224]

[0232] However, the foam is too thick to be fully integrated into at least the EN20 device. , optimal cutting temperature (OCT) After embedding in compound, the specimen was cut on a cryostat to a thickness of approximately 300 microns. Alternatively, silicone foam can also be used in a histology tissue processor. The tissue can also be placed in a container, embedded in paraffin, and sectioned on a microtome. The mixture was removed by multiple washes in water, and the paraffin was removed by subsequent washing in xylene. It is removed by washing repeatedly in alcohol and then in water. Cut the insert to a size that fits inside the device, specifically the EN20 device. Approximately 6 million PECs were then loaded into the EN20 foam device as previously described. The catheter was loaded, sealed, and implanted subcutaneously.

[0225]

[0233] In summary, six EN20 devices were fitted with silicone hollow fiber mats (animal number: 417 4, 4175, 4176, 4177, 4178, and 41879) and porous silicon Each of the 1000 cells was preloaded with a 1000 μg / ml film matrix (data not shown) and an equal number of controls or The EN20 device was not preloaded with matrix (animal number: 4180, 4181, 4182, 4183, 4184, and 4185) (See Figure 72) All 12 devices contain approximately 6 million PECs (the number typically loaded on a device of this size). Immunodeficient mice (e.g., SCID-Beige and Rag) were given approximately twice the amount of IgG1 administered. 2) were implanted subcutaneously on the back of the mice. 13 weeks after implantation, the mice were fasted overnight. Glucose solution was injected intraperitoneally at a dose of 3 mg / kg body weight. Blood samples were taken at fasting time. Serum C-peptides were collected at 10 min and again at 30 and 60 min after glucose administration. The C-peptide was measured using an ELISA kit specific for human C-peptide, and the results are shown in Figure 72. Again, methods for determining serum C-peptide are described in the patent application Serum C-peptide is disclosed ... This has already been amply described in patent and non-patent publications.

[0226]

[0234] The results in Figure 72 show that compared to control animals, the encapsulated PEC implants with silicone hollow fibers: Equivalent function as assessed by serum C-peptide levels at 13 weeks after implantation For example, after 60 minutes of glucose stimulation, Animal number 4174 (1270 pM), derived from a PEC graft with hollow fibers, Serum human C-peptide levels from 75 (977 pM), and 4176 (327 pM) , animal number 4181 (1112 pM), derived from a PEC graft without hollow fibers, and However, none of the PEC hollow fiber implants showed any significant improvement over control animals. The function was not as robust as that observed in the target, 4182.

[0227]

[0235] Animal number 4174 was sacrificed for histological analysis of the PEC hollow fiber implants. Figure 73 shows the hematoxylin and eosin (Figure 73A) and insulin (Figure 73B) Figure 1 shows cross sections of encapsulated grafts or explants stained with PEC. The insulin-positive staining was observed in the hollow fibers. Pigment cells (brown, Figure 73B) are located in the gaps between adjacent hollow fibers (approximately 25 mm from the outer chamber membrane). 6 μm), the silicone fibers act as conduits to allow more This directs oxygen into the core of the chamber. In comparison, cells in the core may become necrotic over time. Although their PEC hollow fiber grafts did not outperform the most robust control PEC grafts, silicone The cellulose-based hollow fibers provide long-term cell viability for these cells in the vicinity of the chamber core. This can bring about advantages.

[0228]

[0236] Example 4 When implanted, the 3D device is inserted (INTERCALLATE) into the body and shaped. Withstands large compressive loads without changing shape One primary embodiment of the present invention is a system capable of high cell volume capacity while at the same time providing a It is a three-dimensional device that, when incorporated into a device, limits the overall surface area or footprint of the device. The three-dimensional large capacity device described herein and described above, when implanted, is completely contained within the body. and maintain their shape and form under various compressive loads. To determine whether the flat (planar) EN250 device The new three-dimensional device was then essentially bent into a three-dimensional bellows-shaped device as described in [74A]. The footprint of the vise is about 50% of the footprint of the original flat sheet device, and the height is The three-dimensional device (Fig. 74A) was implanted in a human cadaver (recently deceased). The subcutaneous fat or skin was then imaged with ultrasound. It is easily inserted into the recess of a 3D device, and the contour of the device is visualized by ultrasound imaging. This indicates that the ion exchange reaction is more easily observed.

[0229]

[0237] The device insertion was successful, so testing was carried out to determine if the device, under high pressure, Will it maintain its curved 3D form or will it flatten and remain in that flattened state? The researchers then determined whether the device would move or migrate from its original implant site. Compressive load was applied to the device implanted subcutaneously (under Scarpia's fascia) Figure 74C shows the same implantation site before application of the compressive load. , shows the same device imaged during and after application of a compressive load. Increasing pressure causes the device to flatten in place, but releasing the load causes the device to retract. The device returns to its original bent bellows shape. The device does not move substantially and the skin does not move. The device remains inserted into the folds and recesses of the chair.

[0230]

[0238] This study demonstrated that the three-dimensional macroencapsulated device described herein, when implanted, , inserted into the body, changing their shape or changing their total footprint. Withstands significant compressive loads without significant migration from the initial implant site This is confirmed.

[0231]

[0239] Example 5 Ultrasound can be used to image the device in vivo A major concern with any cell encapsulation therapy is not only the safety of the cell product, but also , and the safety and integrity of the device. This includes conducting a series of quality control tests (e.g., pressure decay tests) to verify the integrity of the chair. If the encapsulated cell product is implanted in the body, it may cause damage (leakage) of the device. It is possible that a time and event that can cause a device to break down may occur, albeit remotely. , for example, causing the device to expand in a manner inconsistent with normal cell graft production, It may be caused by an internal abnormal cell growth (e.g., cyst, benign tumor). Alternatively, the device may be mechanically implanted at the implant site due to bodily injury or bodily puncture. or physical damage. Therefore, monitoring the device is Visually monitor the device periodically to ensure it remains intact in vivo. It is necessary to check that the item is not damaged.

[0232]

[0240] Therefore, the applicant has developed a simple ultrasound system that is used in many hospitals and doctor's offices. Whether commonly used procedures can be used to monitor implanted devices Ultrasound uses high-frequency sound waves to create images of internal body structures. Ultrasound is non-invasive and does not use radiation, and some ultrasound techniques can produce 3D images. The original image can be created. The ultrasound examination takes 30 or 60 minutes or less. Ultrasound is widely used for examinations. It is widely used and can also reveal dilated blood vessels, blood clots, or narrowed arteries. It can also locate lumps within organs and tissues and can be used to guide needle biopsies. Implantation of the encapsulated cell product is often performed subcutaneously (e.g., in the flank, back, or upper arm). Because the area is expected to be implanted in the area of ​​the ventricle, ultrasound is an easy and convenient outpatient procedure. be.

[0233]

[0241] To investigate the feasibility of ultrasound imaging, an empty flat (planar) EN250 size and EN20 size drug delivery devices and their loaded The device was implanted subcutaneously and imaged. as well as empty (but wetted for imaging purposes) devices. It is also possible to detect loaded devices extended to approximately 1.5 mm and approximately 2.5 mm. In addition, the applicant has already demonstrated that high frequency ultrasound can separate membranes. Not only can the device be detected, but it can also show cyst growth and, with the addition of contrast, can detect blood flow near the device. Therefore, ultrasound imaging can be used to assess the integrity of the device. It is an easy, non-invasive way to regularly monitor the device. If dilation or device breakage is observed, the device should again be removed from the outpatient The procedure involves surgically removing it from the body.

[0234]

[0242] Examples 4 and 5 show the device being used for in vivo imaging and monitoring. This document demonstrates that no invention is necessary to achieve the objectives of in vi Imaging and external testing of device integrity in vo, these means and other means are envisioned.

[0235]

[0243] Accordingly, it is understood that those skilled in the art will be able to readily adapt the present invention to the present application without departing from the scope and spirit of the present invention. Various substitutions, modifications or optimizations, or combinations may be made to the embodiments disclosed herein. It is clear that

[0236]

[0244] As used in the claims below and throughout this disclosure, "consisting essentially of" The phrase includes any element listed after the phrase, but does not include the listed elements. , other compounds that do not interfere with or contribute to the activity or action specified in this disclosure. It means that no element is excluded. Thus, the phrase "consisting essentially of" means that The listed elements are required or mandatory, while other elements are optional and These may be present or absent depending on whether they affect the activity or action of the listed elements. It indicates that something does not have to exist.

[0237]

[0245] (Embodiment) Embodiment 1: A cell encapsulation assembly for implantation into a mammalian recipient, comprising: at least two chambers for enclosing cells, and a first seal portion, thereby forming an enclosed assembly; At least a second seal is located within the cell containment assembly and forms the inner periphery of the cell chamber. A cell encapsulation assembly comprising:

[0238]

[0246] Embodiment 2: The second seal portion is further folded at an angle to form an assembly. Reduced total footprint compared to an assembly without a fold in the second seal 2. The assembly of embodiment 1,

[0239]

[0247] Embodiment 3: With or without folds in the second seal , maintaining substantially the same cell volume capacity.

[0240]

[0248] Embodiment 4: The assembly of embodiment 1, comprising a semipermeable membrane.

[0241]

[0249] Embodiment 5: A third seal or a second seal is inserted into the cell chamber formed by the second seal. 10. The assembly of claim 1, having a first or fourth seal.

[0242]

[0250] Embodiment 6: The method of embodiment 1, further comprising at least one loading port. Assembly.

[0243]

[0251] Embodiment 7: The assembly of embodiment 1, further comprising two loading ports. -.

[0244]

[0252] Embodiment 8: The assembly of embodiment 1, further comprising living cells.

[0245]

[0253] Embodiment 9: The live cells are human PDX1 (pancreatic and duodenal leukemia) cells. 10. The method of claim 6, wherein the cells are human homeobox gene 1)-positive pancreatic progenitor cells. Assembly.

[0246]

[0254] Embodiment 10: The assembly of embodiment 6, wherein the live cells are human endocrine precursor cells. -.

[0247]

[0255] Embodiment 11: A first sealed periphery and an inner periphery seal of the cell chamber. and a second seal portion within the first seal portion, forming a second seal portion within the cell chamber. the device assembly, wherein the sealing portion and the bulkhead seal portion do not increase the surface area of ​​the device assembly. Mumble.

[0248]

[0256] Embodiment 12: The assembly of embodiment 11, wherein the second seal is folded. Lee.

[0249]

[0257] Embodiment 13: As described in embodiment 12, wherein the second seal portion is folded 0 to 90 degrees. Assembly of.

[0250]

[0258] Embodiment 14: As described in embodiment 12, wherein the second seal portion is folded 0 to 45 degrees. Assembly of.

[0251]

[0259] Embodiment 15: As described in embodiment 12, wherein the second seal portion is folded 0 to 30 degrees. Assembly of.

[0252]

[0260] Embodiment 16: As described in embodiment 12, wherein the second seal portion is folded 0 to 40 degrees. Assembly of.

[0253]

[0261] Embodiment 17: As described in embodiment 12, wherein the second seal portion is folded 0 to 90 degrees. Assembly of.

[0254]

[0262] Embodiment 18: The assembly of embodiment 12, wherein the second seal is folded 0 degrees. Mumble.

[0255]

[0263] Embodiment 19: The adhesive of embodiment 12, wherein the second seal is folded 40 degrees. Centrifuge.

[0256]

[0264] Embodiment 20: The second seal portion, when folded, reduces the total footprint of the assembly. 13. The assembly of embodiment 12, wherein

[0257]

[0265] Embodiment 21: The device further includes a septum seal, the septum seal being within the second seal. 12. The assembly of embodiment 11, wherein the thickness of the chamber is reduced.

[0258]

[0266] Embodiment 22: The method of embodiment 11, further comprising at least one loading port. Assembly shown.

[0259]

[0267] Embodiment 23: The assembly of embodiment 11, further comprising two loading ports. Bree.

[0260]

[0268] Embodiment 24: The assembly of embodiment 11, further comprising living cells.

[0261]

[0269] Embodiment 25: The live cells are human PDX1 (pancreatic and duodenal leukemia) cells. nal homeobox gene 1)-positive pancreatic progenitor cells, Assembly as shown.

[0262]

[0270] Embodiment 26: The assembly of embodiment 24, wherein the live cells are human endocrine precursor cells. Lee.

[0263]

[0271] Embodiment 27: The method of embodiment 11, further comprising a matrix inside the chamber. Centrifuge.

[0264]

[0272] Embodiment 28: The matrix is ​​a biostable material that facilitates oxygen and nutrient uptake. 28. The assembly of embodiment 27, comprising a material.

[0265]

[0273] Embodiment 29: A medical device configured to reduce the footprint of a device. Medical device.

[0266]

[0274] Embodiment 30: A device assembly comprising at least two cell chambers.

[0267]

[0275] Embodiment 31: Two chambers in a first unfolded configuration. 31. The assembly of embodiment 30.

[0268]

[0276] Embodiment 32: The two chambers are in a second fold configuration. 32. The assembly of embodiment 31.

[0269]

[0277] Embodiment 33: A device assembly comprising at least two chambers, Chambers are configured to reduce the footprint of the device assembly Device assembly.

[0270]

[0278] Embodiment 34: Configurable to reduce footprint of device assembly 34. The method of claim 33, wherein the surface area of ​​the chamber maintains the same surface area when the chamber is moved. Assembly.

[0271]

[0279] Embodiment 35: At least two cell chambers for encapsulating living cells, cell chambers It contains acellular regions along the long axis separating the bars, which bend to form folds. ,The folds compare the effective area of ​​the assembly with the assembly without folds. and reducing the number of cells in the cell encapsulation assembly, thereby forming a three-dimensional cell encapsulation device. Lee.

[0272]

[0280] Embodiment 36: Substantially the same details with and without folds 36. The assembly of embodiment 35, which maintains cell volume capacity.

[0273]

[0281] Embodiment 37: The assembly of embodiment 35, comprising a semipermeable membrane.

[0274]

[0282] Embodiment 38: At least two, three, four, five, six, seven, eight or more cell channels. 36. The assembly of embodiment 35, comprising a member.

[0275]

[0283] Embodiment 39: The method of embodiment 35, further comprising at least one loading port. Assembly as shown.

[0276]

[0284] Embodiment 40: The assembly of embodiment 35, further comprising two loading ports. Bree.

[0277]

[0285] Embodiment 41: The assembly of embodiment 35, wherein the living cells are definitive endoderm lineage cells. Lee.

[0278]

[0286] Embodiment 42: The live cells are human PDX1 (pancreatic and duodenal leukemia virus) nal homeobox gene 1)-positive pancreatic progenitor cells, Assembly as shown.

[0279]

[0287] Embodiment 43: The assembly of embodiment 35, wherein the live cells are human endocrine precursor cells. Lee.

[0280]

[0288] Embodiment 44: The acetonide of embodiment 35, wherein the live cells are human immature beta cells. Bree.

[0281]

[0289] Embodiment 45: Multiple microorganisms capable of dispersing live cells and improving oxygen distribution inside the cell chamber. The embodiment further comprises a cell chamber matrix having interconnected cavities or pores. 36. The assembly of embodiment 35.

[0282]

[0290] Embodiment 46: The assembly of embodiment 45, wherein the interconnected cavities have different cavity dimensions. Bree.

[0283]

[0291] Embodiment 47: The matrix is ​​polydimethylsiloxane (PDMS), polydimethyl Siloxane monoacrylate, and polydimethylsiloxane monomethacrylate 46. ​​The assembly of embodiment 45.

[0284]

[0292] Embodiment 48: As described in embodiment 45, wherein the matrix is ​​a silicone elastomer. Assembly of.

[0285]

[0293] Embodiment 49: An embodiment in which the matrix is ​​polydimethylsiloxane (PDMS) 46. ​​The assembly according to claim 45.

[0286]

[0294] Embodiment 50: The method of embodiment 35, wherein the cell chambers are parallel to each other. Centrifuge.

[0287]

[0295] Embodiment 51: The method of embodiment 35, wherein the cell chambers are separated by about 20 degrees. Centrifuge.

[0288]

[0296] Embodiment 52: The method of embodiment 35, wherein the cell chambers are separated by about 40 degrees. Centrifuge.

[0289]

[0297] Embodiment 53: The method of embodiment 35, further comprising a septum seal within the cell chamber. Assembly.

Claims

1. A cell encapsulation assembly, comprising: at least two substantially oval or rectangular shaped cell chambers and a cell-free region along a longitudinal axis of the cell containment assembly separating the at least two cell chambers; the cell-free region bends to form a fold; each cell chamber contains an in vitro viable cell population, said viable cell population comprising definitive endoderm-lineage cells, human immature beta cells or endocrine precursor cells; the cell chamber is porous, allowing molecules produced by the cells to diffuse from the cell chamber into the surrounding tissue; Cell encapsulation assembly.

2. A cell encapsulation assembly, comprising: at least two substantially oval or rectangular shaped cell chambers and a cell-free region along a longitudinal axis of the cell containment assembly separating the at least two cell chambers; the cell-free region bends to form a fold; each cell chamber contains an in vitro viable cell population, said viable cell population comprising definitive endoderm-lineage cells, human immature beta cells or endocrine precursor cells; the cell chamber is bounded by a semipermeable membrane; Cell encapsulation assembly.

3. 3. The assembly of claim 1 or 2, further comprising two loading ports for loading cells into the cell chamber.

4. The assembly of claim 3 , wherein the loading port comprises a tube.

5. 5. The assembly of any one of claims 1 to 4, wherein the assembly can be sealed once cells are loaded into the assembly.

6. 6. The assembly of any one of claims 1 to 5, wherein the folds reduce the effective area of ​​the assembly compared to an assembly without the folds, thereby forming a three-dimensional cell-encapsulating assembly.

7. 7. The assembly of any one of claims 1 to 6, comprising at least 3, 4, 5, 6, 7, 8 or more cell chambers.

8. The assembly of any one of claims 1 to 7, wherein the cell chambers are parallel to each other.

9. 9. The assembly of claim 1, further comprising a cell chamber matrix within each cell chamber, the cell chamber matrix having a plurality of interconnected cavities or pores, which disperses live cells loaded within the cell chambers and improves oxygen distribution within the cell chambers.

10. The assembly of any one of claims 1 to 9, wherein the live cell population comprises definitive endoderm lineage cells or human immature beta cells.

11. The assembly of claim 10 , wherein the viable cell population comprises definitive endoderm lineage cells.

12. 11. The assembly of claim 10, wherein the population of live cells comprises human immature beta cells.

13. The assembly of any one of claims 1 to 9, wherein the population of live cells comprises endocrine precursor cells.

14. 14. The assembly of claim 1, wherein the cell-free region is bent at an angle of between 5 degrees and 175 degrees.

15. 15. The assembly of any one of claims 1 to 14, wherein the assembly comprises at least three cell chambers and a folded cell-free region separating adjacent cell chambers.

Images