Vascularized device and method for islet transplantation without chronic immunosuppression

Abstract

Disclosed herein are systems, devices, and method for implanting biological material in a subject in need thereof. The devices comprise a non-mechanically immune-isolating space defined by a protective, perforated frame and a placeholder disposed within the frame. The frame and plunger allow for vascularization to occur through the device. The plunger may be removed and therapeutic biological material comprised of a carrier matrix, tolerance inducing materials and therapeutic cells are deposited in the volume vacated by the plunger. The systems, devices, and methods allow for therapeutic deposition of transplanted cells without the need for long-term global immunosuppression.

Description

VASCULARIZED DEVICE AND METHOD FOR ISLET TRANSPLANTATIONWITHOUT CHRONIC IMMUNOSUPPRESSIONCROSS-REFERENCE TO RELATED APPLICATION(S)

[0001] This application claims priority to and any benefit of U.S. Provisional Application No. 63 / 647,556, filed May 14, 2024, the content of which is incorporated herein by reference in its entirety.FIELD

[0002] The present invention relates to apparatuses, systems, and methods for use in therapeutic tissue implantation.BACKGROUND

[0003] Diabetes is a chronic condition where the body either doesn’t produce enough insulin or can’t effectively use the insulin it produces, leading to high blood sugar levels which can lead to severe symptoms, both chronic and acute. There are three main types of diabetes, Type 1, Type 2, and gestational diabetes. Type 1 diabetes (T1D) is a chronic disease where the body's immune system mistakenly attacks and destroys the insulin-producing cells in the pancreas. This leads to the body not producing enough insulin, or not producing any at all. Without insulin, the body can't properly use glucose for energy, causing blood sugar levels to rise. This can lead to serious health problems or even death if it’s not treated. People with Type 1 diabetes need synthetic insulin every day in order to live and be healthy. Type one diabetes affects nearly 1.25 million Americans.

[0004] Cell replacement therapy is a promising potential treatment option for a wide variety of diseases, including T1D. Many clinical conditions and disease states result from the lack of factor(s) produced by living cells or tissues, including, for example, diabetes, in which insulin production is inadequate; Parkinson's disease, in which dopamine production is decreased; and anemia, in which erythropoietin is deficient. Such conditions or diseases may be treated by cell / tissue implants that produce the missing or deficient factor(s).

[0005] Human tissue / cell transplantation has been successfully performed for decades. While advances have been made in the field, many challenges remain for successful cell replacement therapy. The viability and functionality of transplanted cells can be compromised by, for example, lack of mechanical protection of the cells, lack of necessary factors for growth and establishment of the cells in the body, e.g. by inadequate vascularization or by inability of thevascular system to reach parts of the transplant, and anti-transplant host immune activity. Importantly, conventional cell and tissue transplanting methods require immune suppression that carries with it a host of problems for the recipient.

[0006] Attempts have been made to provide devices that mechanically isolate cells during transplantation to facilitate engraftment. See, e.g., U.S. Pat. No. 8,702,684. Devices such as these often require complex drug-delivery apparatuses connected to the cell compartment for delivery of immunomodulatory compounds. The devices also required reloading and other challenges related to connectivity between the portions of the device. These drawbacks led to a lack of success in pre-clinical testing that did not warrant to proceed for clinical trials.SUMMARY

[0007] The general inventive concepts are based, in part, on the recognition that conventional apparatuses, systems, and methods for tissue / cell transplantation are ineffective in the long term for one or more reasons related to vascularization and transplant rejection. The general inventive concepts address these issues with an innovative device designed to promote rapid, stable vascularization. The device allows for efficient tissue transfer for implantation and engraftment of cells / tissues, with a single device that can house a predetermined volume (e.g., approximately 4-15 ml in the clinical relevant models, while for pre-clinical testing there are devices made between 150, 450 and 1,500 microliters, for rodent and large animal research applications, respectively).

[0008] Despite significant progress, the clinical large scale applicability of cell replacement strategies remains severely limited by the requirement of recipient immunosuppression which imposes a risk of severe adverse events and long-term transplant failure. The general inventive concepts address the problem of transplant rejection by providing localized tolerance induction (immunosuppression / immunoregulation) at the site of cellular transplantation, while avoiding the requirement for long term systemic immunosuppression of the subject. The invention also addresses these problems by providing local delivery of factors beneficial to transplanted tissue / cells, such as, e.g., those that favor cell engraftment, growth, and function.

[0009] The general inventive concepts provide a device in the form of a scaffold to promote vascularization while also providing mechanical protection. Once vascularization is underway, the device provides mechanical protection for both a tolerance-inducing drug delivery system and a therapeutic cell product (e.g., allogenic pancreatic islets) and addresses many of the drawbacks of conventional systems and methods for cell / tissue transplantation. The systemsand methods of the general inventive concepts enable biologic cures without the need for global immunosuppression (IS), by providing a system combining 1) a vascularizing device that allows for blood oxygen / nutrient delivery to the implanted tissues / cells, 2) a carrier matrix (e.g., microfragmented fat tissue) for prolonged (e.g., longer than 1 week) local delivery of tolerance inducing drugs, cells or other biologies that can be preloaded into the carrier matrix, and 3) a therapeutic cells / tissue (e.g., an islet product). In certain embodiments, the system also comprises a tolerance-inducing material including but not limited to SA-FasL and anti-CD40L, including a soluble protein form of SA-FasL, which may also be incorporated or otherwise preloaded into the MF AT with the therapeutic cell / tissue product.

[0010] In certain embodiments, the biological material is a composition comprising one or more select cell types or tissues. In certain of these embodiments, the general inventive concepts further address the problem of rejection of the therapeutic cells by providing a carrier matrix and incorporating / loading the cells within the carrier matrix, which provides protection from mechanical stress and from the host immune system. In certain exemplary embodiments, incorporating the cells in the carrier matrix may reduce or preferably minimize the need for even local delivery of immunosuppressive / immunoregulatory substances. The cells are further protected by the mechanical support provided by the device.

[0011] In a broad sense, the general inventive concepts provide a device for rapid vascularization comprising a perforated container vessel (i.e., a mechanoprotective frame or shell) containing a plunger, which when engaged in the frame forms a gap of 0.5 mm - 2 mm between the frame and the plunger to allow for vascularization to occur; the plunger contains one or more hollow pass-through conduits (e.g., a through-hole) to allow for loading of carrier matrix and tissue / cell products while withdrawing the plunger, after vascularization has occurred.

[0012] In certain exemplary embodiments, the general inventive concepts contemplate a system for the implantation of therapeutic tissues. The system comprises a device including a perforated mechanoprotective frame defining an inner space for receiving biological material, the perforated mechanoprotective frame comprising a first opening and a second opening opposing the first opening, a plunger positioned in the inner space of the frame and which is removable before deposition of the biological material, an outer surface of the plunger defining a vascularization volume with an inner surface of the perforated mechanoprotective frame, and a cap positioned in a first opening in the perforated mechanoprotective frame opposite to theplunger; and biological material comprising a therapeutic cell, a carrier matrix, and a tolerance inducing drug, cell, or other biologic.

[0013] In certain exemplary embodiments, the general inventive concepts contemplate a method of implanting biological material in a patient. The method comprises implanting into the patient a device comprising a perforated mechanoprotective frame defining an inner space for receiving biological material, the perforated mechanoprotective frame comprising a first opening and a second opening opposing the first opening, a plunger positioned in the inner space of the frame and which is removable before deposition of the biological material, an outer surface of the plunger defining a vascularization volume with an inner surface of the perforated mechanoprotective frame, and a cap positioned in a first opening in the perforated mechanoprotective frame opposite to the plunger; allowing tissue ingrowth into and / or around the inner space of the device, accessing the device and removing the plunger element and depositing the biological material into the space vacated by the plunger element, the biological material comprising a therapeutic cell, a carrier matrix, and a tolerance inducing drug, cell, or other biologic; and positioning a cap element in an open end of the device.

[0014] Other aspects and features of the general inventive concepts will become more readily apparent to those of ordinary skill in the art upon review of the following description of various exemplary embodiments in conjunction with the accompanying figures.BRIEF DESCRIPTION OF THE DRAWINGS

[0015] These and other features of the present disclosure will become better understood with regard to the following description and accompanying drawings in which:

[0016] Figure 1 shows top and bottom views of three exemplary devices according to the general inventive concepts.

[0017] Figure 2 shows top and bottom views of the devices shown in Figure 1 with additional front views (along the bottom of the drawing) and cross-sectional views (along the top of the drawing) of each device.

[0018] Figure 3 is a perspective view of an embodiment of a mechanoprotective frame according to the general inventive concepts with an enhanced view of the mesh wall of the frame.

[0019] Figure 4 is a perspective view of a device according to the general inventive concepts.

[0020] Figure 5A is a top view of a device according to the general inventive concepts wherein the plunger is partially withdrawn from the frame.

[0021] Figure 5B is a perspective view of a device according to the general inventive concepts wherein the plunger is partially withdrawn from the frame.

[0022] Figure 6A is a top view of a device according to the general inventive concepts.

[0023] Figure 6B is a perspective view of a device according to the general inventive concepts.

[0024] Figure 6C is a front view of a device according to the general inventive concepts.

[0025] Figure 6D is a back view of a device according to the general inventive concepts.

[0026] Figure 6E is a side cross-sectional view of a device according to the general inventive concepts.

[0027] Figure 6F is a front cross-sectional view of a device according to the general inventive concepts.

[0028] Figure 7A shows a top view of a mechanoprotective frame according to the general inventive concepts.

[0029] Figure 7B is a side cross-sectional view of the mechanoprotective frame of Figure 3A.

[0030] Figure 7C is a front view of the mechanoprotective frame of Figure 3 A.

[0031] Figure 7D is a back view of the mechanoprotective frame of Figure 3 A, showing the cap inserted into the top opening.

[0032] Figure 8A is a perspective view of a cap for the mechanoprotective frame according to the general inventive concepts.

[0033] Figure 8B is a bottom view of the cap.

[0034] Figure 8C is a back view of the cap.

[0035] Figure 8D is a cross-sectional view of the cap.

[0036] Figure 9A is a top view of an embodiment of a device according to the general inventive concepts.

[0037] Figure 9B is a perspective view of an embodiment of a device according to the general inventive concepts.

[0038] Figure 9C is a back view of an embodiment of a device according to the general inventive concepts.

[0039] Figure 9D a side cross-sectional view of an embodiment of a device according to the general inventive concepts.

[0040] Figure 9E is a bottom view of an embodiment of a device according to the general inventive concepts.

[0041] Figure 10A is a top view of a mechanoprotective frame for a device according to the general inventive concepts.

[0042] Figure 10B is a perspective view of a mechanoprotective frame according to the general inventive concepts.

[0043] Figure 10C is a back view of a mechanoprotective frame according to the general inventive concepts.

[0044] Figure 10D is a front view of a mechanoprotective frame according to the general inventive concepts.

[0045] Figure 10E is a side cross-sectional view of a mechanoprotective frame according to the general inventive concepts.

[0046] Figure 10F an enhanced view of an embodiment of the mesh wall of the mechanoprotective frame.

[0047] Figure 11 is a back perspective view of a plunger according to the general inventive concepts.

[0048] Figure 12A is a top view of a plunger according to the general inventive concepts.

[0049] Figure 12B is a cross-sectional view of the plunger.

[0050] Figure 13 A is a back view of the plunger (i.e., from the perspective of the insertion end).

[0051] Figure 13B is a front view of the plunger (i.e., from the perspective of the cap end).

[0052] Figure 14 is an image of a device according to the general inventive concepts.

[0053] Figure 15A is an image of a disassembled device according to the general inventive concepts.

[0054] Figure 15B is an image showing an assembled device according to the general inventive concepts.

[0055] Figure 16A is a perspective view of an alternative embodiment of a plunger.

[0056] Figure 16B is a front view of the plunger shown in 16A.

[0057] Figure 17A is a top view of the plunger shown in 8A.

[0058] Figure 17B is a cross-sectional view of the plunger shown in 8 A.DETAILED DESCRIPTION

[0059] Despite significant progress, wide-scale applicability of islet transplantation for e.g., diabetes (or other disease modifying cell therapies), remains limited by the requirement of lifelong recipient immunosuppression (IS), imposing long-term risks of adverse events and transplant failure.

[0060] Various technologies pertaining to an apparatus, system, and method for cell / tissue transplantation in subjects suffering from diabetes mellitus, including T1D, and related conditions, are described herein. The general inventive concepts provide a method to use transplanted cells for alleviation and / or treatment of the conditions or diseases, or symptoms thereof, described herein.

[0061] The general inventive concepts relate to systems for and methods of implanting tissue in a subject. In a broad sense, the general inventive concepts improve on conventional tissue implanting devices and methods by providing means for rapid, stable vascularization of animplantation site, while simultaneously providing a carrier matrix (e.g., a drug delivery biomaterial) for supporting / protecting the transplanted cells and transient immunomodulation while tolerance induction takes place.

[0062] The terminology as set forth herein is for description of the embodiments only and should not be construed as limiting the disclosure as a whole. All references to singular characteristics or limitations of the present disclosure shall include the corresponding plural characteristic or limitation, and vice versa, unless otherwise specified or clearly implied to the contrary by the context in which the reference is made. Unless otherwise specified, “a,” “an,” “the,” and “at least one” are used interchangeably. Furthermore, as used in the description and the appended claims, the singular forms “a,” “an,” and “the” are inclusive of their plural forms, unless the context clearly indicates otherwise.

[0063] As used herein, the term “or” is intended to mean an inclusive “or” rather than an exclusive “or.” That is, unless specified otherwise, or clear from the context, the phrase “X employs A or B” is intended to mean any of the natural inclusive permutations. That is, the phrase “X employs A or B” is satisfied by any of the following instances: X employs A; X employs B; or X employs both A and B. In addition, the articles “a” and “an” as used in this application and the appended claims should generally be construed to mean “one or more” unless specified otherwise or clear from the context to be directed to a singular form.

[0064] Ranges as used herein are intended to include every number and subset of numbers within that range, whether specifically disclosed or not. Further, these numerical ranges should be construed as providing support for a claim directed to any number or subset of numbers in that range. For example, a disclosure of from 1 to 10 should be construed as supporting a range of from 2 to 8, from 3 to 7, from 5 to 6, from 1 to 9, from 3.6 to 4.6, from 3.5 to 9.9, and so forth.

[0065] Any combination of method or process steps as used herein may be performed in any order, unless otherwise specified or clearly implied to the contrary by the context in which the referenced combination is made.

[0066] The terms “modulating” or “modulation” or “modulate” as used herein, unless otherwise specified, refer to the targeted movement of a selected characteristic (e.g., insulin level, blood sugar level, HbAlc level, macrophages, regulatory or effector T Cells)

[0067] The term “ameliorate” as used herein, unless otherwise specified, means to eliminate, delay, or reduce the prevalence or severity of symptoms associated with a condition.

[0068] The term “an effective amount” as used herein, unless otherwise specified, is intended to qualify the amount of a composition according to the general inventive concepts (e.g., therapeutic cells or cellular products thereof, pancreatic islets, SA-FasL, insulin, or related cellular products) which will achieve the goal of decreasing the risk that the individual will suffer an adverse health event (e.g., rejection of implanted tissue / cell or an event related to high blood sugar level), including reducing one or more symptoms, while avoiding adverse side effects such as those typically associated with alternative therapies (e.g., chronic recipient immunosuppression).

[0069] The terms “treating”, and “treatment” as used herein, unless otherwise specified, includes delaying the onset of a condition, reducing the severity of symptoms of a condition, or eliminating some or all of the symptoms of a condition.

[0070] The terms “therapeutic cells,” “implanted cells,” and “transplanted cells” as used herein, unless otherwise specified, refer to cells that provide one or more targeted biologic factors or hormones to an individual that modulate a symptom experienced by the individual. In certain exemplary embodiments, the cells are selected from pancreatic islets, adipose tissue products, mesenchymal stromal cells, and could include subcellular products such as exosomes and extracellular vesicles (EVs).

[0071] The term “carrier matrix” or “biological carrier matrix” as used herein, unless otherwise specified, refers to a biologically derived drug delivery product such as, but not limited to micro-fragmented adipose tissue (MFAT), which is adipose tissue that is isolated from an animal (e.g., a human, including but not limited to the subject) and is processed according to microfat processing methods or devices. One suitable procedure for obtaining MFAT is disclosed in the patent application WO2011 / 145075, the content of which is incorporated by reference. The fat tissue, preferably the lipoaspirate, is introduced in a Lipogems® device wherein it is progressively reduced (fragmented) in small clusters of fat tissue preferably by means of mild mechanical forces and, more preferably, in presence of a solution, preferably a saline solution. According to a preferred embodiment, the microfragmented fat of the invention contains clusters of fat tissue that are characterized by retaining the natural / intact stromal vascular niche of the resident cells that, consequently, are supportedby the stroma resembling the natural / physiological context in trophic and / or signaling terms. Additionally, the stroma provides a protected environment during the graft of the cells against many physical and / or chemical insults.

[0072] The term “tolerance inducing drugs, cells, or other biologies” as used herein, unless otherwise specified, refers to therapeutic products that induce local tolerance to aid in transplantation by slowing or modulating the immune “non self’ response, facilitating enhanced implantation outcomes. Examples of suitable tolerance inducing materials include known immunosuppressants such as rapamycin or anti-CD40L antibodies, PD-L1, SA FasL or other FasL soluble or oligomeric formulations. In certain embodiments, the tolerance inducing drugs, cells or other biologies is a tolerance inducing biologic such as streptavidin FasL. In addition the MFAT could also provide benefits related to its anti-inflammatory, immunomodulatory, and protection against ischemia-revascularization injury.

[0073] To achieve the foregoing and other objects, the general inventive concepts provide a device that enables cellular therapy to be performed by implantation into a subject in need thereof. In certain exemplary embodiments, the general inventive concepts encompass combining the vascularizing device with a carrier matrix (e.g., microfragmented fat tissue) for prolonged (>1 week) local delivery of tolerance inducing drugs, cells or other biologies that can be preloaded into the carrier matrix (e.g., delivery tissue), as exemplified herein. In certain embodiments, the drug delivery tissue or biomaterial (e.g., FasL, anti-CD40L, MSC, Exosomes and EVs), can also confer protection from ischemia-revascularization injury (e.g., micro fragmented fat / stromal vascular fraction / pericytes, MSC / endothelial cells), while new blood vessels re-populate the inner portion of the device (cell / tissue compartment).

[0074] In certain embodiments, the general inventive concepts provides a device that comprises a microenvironment favorable to cell and / or tissue survival and function, e.g. by providing a vascularized bed for the implanted biological material; and a carrier matrix that may include one or more nutrients, factors, cytokines, and immunosuppressive / immunoregulatory molecules facilitating effective and stable implantation of the tissue / cells.

[0075] In certain embodiments, some or all of the implanted biological material (e.g. a composition comprising one or more types of cells, tissues, or cell products, or a combination thereof) is enclosed / incorporated within in a biocompatible carrier matrix. The cells areprotected from physical trauma by the mechanical support provided by the device.

[0076] In certain exemplary embodiments, the general inventive concepts are embodied in a device, which may be implanted and vascularized, for receiving implanted biological material (i.e., therapeutic tissue / cells), which may be encapsulated in a carrier matrix, comprising: a surface defining and mechanically shielding an adjacent space, such as an inner space / volume. The device may be of any shape suitable for implantation and may vary based on the biological material to be implanted, the intended therapeutic effect, and / or the location of the implant, for example. Those of ordinary skill in the art can assess the shape(s) preferred for the intended application(s). In certain embodiments, the device is a cage-like device, as shown and described herein, in which new capillaries grow through a mechanoprotective mesh / frame.

[0077] Some features of the device according to the general inventive concepts include the following: the vascularizing device for islet transplantation (VADIT) is designed to provide rapid implantation, vascularization, and engraftment of cells / tissues. In the clinically relevant model, the device can house approximately 4-15 ml of cell / tissue product. The VADIT device is not mechanically immune-isolating and is made up of a hollow, rounded rectangular protective frame or shell with a closed end and a removeable plunger / placeholder that mates with the interior of the frame or shell, leaving a space or vascularization depth of approximately 1 mm (0.5 mm to 1.5 mm) on all sides between the outer surface of the plunger (114c) and the inner surface of the frame or shell, for vascularization (i.e., defining a vascularization volume). The frame has open ends and is made of a perforated medical-grade stainless steel, titanium, pique, or other medical grade metal or plastic material. The hexagonal or circular perforations of the shell are sized to promote vascularization, wherein the holes are approximately 0.4 mm (400 microns). The walls of the shell are approximately 0.5 mm in thickness. Note: The overall lengthwise and widthwise dimensions will be proportionally increased depending on the size of the recipient animal or human application. The only dimension held constant is the thickness of the cell / tissue product compartment that must remain approximately 3mm.

[0078] In certain embodiments, the device comprises an outer flask-shaped frame / shell enclosing an adjacent, e.g. inner, space or cavity. In certain embodiments, the device may contain a plunger that is positioned within the frame. After a period of time that allows for adequate vascularization of the area in and around the device, the plunger may be removed, and the space formerly occupied by the plunger may be filled with the biological material (i.e., therapeutic cells) to be transplanted.

[0079] Turning to the drawings, Figure 1 shows three exemplary embodiments of a device 100 according to the general inventive concepts. The device is a substantially flat, rounded rectangle having a length and width defining first and second major surfaces and a thickness extending therebetween. As can be seen from the figure, the length and width may vary substantially based on the size / species of recipient, with larger species (e.g., human) requiring a larger device. The general inventive concepts are based, in part, on the recognition that a minimum thickness of the device (e.g., about 0.4 cm to about 1 cm, including about 0.5 cm to about 0.7 cm, and including about 0.6 cm) is required to facilitate suitable vascularization in the device. The device comprises a mechanoprotective frame 110 formed from a durable biocompatible mesh (e.g., stainless steel, titanium, or biocompatible plastic material), defining a cavity within. In certain embodiments, the device further comprises a cap 116 on at least one opening end of the frame. In certain exemplary embodiments, the overall thickness of the device / frame is about 0.4 cm to about 1.0 cm, including, about 0.6 cm or less. This thickness provides a 3 mm thickness of the tissue / cell product volume (placeholder / plunger component) and 1 mm all around the plunger (when engaged / positioned within the frame) for vascularization in the volume between the frame and plunger. In certain embodiments, the device further comprises a plunger 112 or placeholder positioned in an end of the frame. The plunger may include one or more (in the clinically relevant device) through-holes 113, which are channels formed through the length of the plunger for delivery of materials to the interior of the device. In each of the three embodiments shown in Figure 1, the right-most figure of each pair of images represents an embodiment of the device including a plunger / pl aceholder and the left of the two images shows an embodiment of the device with a cap that is positioned in the opening after removal of the placeholder and after introducing the biological material.

[0080] Figure 2 shows the three devices shown in Figure 1 with additional front views along the lower portion of the drawing and corresponding cross-sectional views along the top of the drawing. The through-hole is visible on the right-most of each pair of embodiments.

[0081] Figure 3 shows an embodiment of a device 100 according to the general inventive concepts. As can be seen from the figure, the frame has a generally rounded rectangular shape with openings 120a and 120b, that have a generally elliptical shape. The device is formed from a durable biocompatible mesh (e.g., the mechanoprotective frame or mesh can be of stainless steel, polymer or any other suitable material that will provide dimensional stability), to provide a mechanoprotective frame, defining an adjacent, e.g., inner, space or cavity 115. Themechanoprotective frame is perforated sufficiently so as to permit capillaries to grow through the perforations to provide a vascular bed for promoting engraftment of transplanted cells. The perforations in the frame may be, e.g., about 100-1000 microns, including about 200-800 microns, including about 300-600 microns, and including about 350-500 microns. By way of example, a stainless steel mesh with holes / perforations of about 400 microns (diameter) may be provided, but the holes could be slightly smaller or bigger depending on the therapeutic needs among other factors. Any other size that permits adequate vascularization for the specific device location and therapeutic regime is envisioned as being part of the present invention. The device houses a therapeutic biological material, e.g., cells / tissue or products thereof, either at the time of implantation or in a second stage (after vascularization of the device). Inn certain embodiments, the device and / or plunger further comprise a tether or some mechanism for retrieval and cap placement.

[0082] Figure 4 is a perspective view of an embodiment of a device 100 according to the general inventive concepts. The device comprises a mechanoprotective frame 110, a plunger / pl aceholder 112, which includes a through -hole 113 (shown formed through the length of the plunger, for delivery of materials to the interior of the device). When inserted, the plunger is positioned in the inner space of the frame and is removable before deposition of the biological material. The outer surface of the plunger (that which is within the frame) defines a vascularization volume with the inner surface of the perforated mechanoprotective frame.

[0083] Figure 5 A shows a top view and figure 5B shows a perspective view of an embodiment of a device 100 according to the general inventive concepts. The device comprises a mechanoprotective frame 110 and a plunger / pl aceholder 112, which includes a through-hole. The plunger is shown partially withdrawn or removed from the frame, showing the positioning during use of the device upon vascularization wherein the biological materials would be introduced into the adjacent space held by the plunger.

[0084] Figure 6A is a top view of a device 100 according to the general inventive concepts. The device has a length 1, a width w, and a thickness t. In certain embodiments, the thickness of the device remains consistent between the various embodiments shown herein, whereas the length and width may vary (corresponding to the size of the respective subject for which it is intended). In certain embodiments, the thickness is approximately 3mm in the cavity for tissue / cell product volume (placeholder / plunger component). This allows for approximately 1mm all around the plunger for vascularization to occur between the frame and plunger. Figure6B is a perspective view of the device. Figure 6C is a front view of the device. Figure 6D is a back view of the device. Figure 6E is a side cross-sectional view of the device. Figure 6F is a front cross-sectional view of the device.

[0085] Figure 7A shows a top view of the mechanoprotective frame shown in Figure 3 with a cap 116 inserted in a first end of the frame and an open end 120b opposite the cap. In embodiments wherein the plunger is positioned in the frame, the cap is positioned in the opening opposite o that of the plunger. Figure 7B is a side cross-sectional view of the mechanoprotective frame of Figure 7A. Figure 7C is a front view of the mechanoprotective frame of Figure 3A, which shows the elliptical shape of the opening and the frame itself. The mechanoprotective frame preferably has rounded edges so as to be relatively ergonomic, to be comfortable to the subject while implanted, and to minimize stress and / or injury to the subject. Figure 7 is a back view of the mechanoprotective frame of Figure 7A, showing the cap positioned into an opening. In certain embodiments, the cap is removably positioned in the opening, in alternative embodiments, the cap is not removeable from the frame.

[0086] During the vascularization phase, one end of the cavity 115 is closed with the head or cap end 114a of the plunger 112 that is selectively disposed within the cavity to define the volume for the new capillaries. In certain exemplary embodiments, the cap of the plunger is sized and configured to mate with an opening 120a / 120b of the frame during vascularization, similar to that of the cap. The opposite opening of the frame is closed with a cap 116.

[0087] Figure 8 A is a perspective view of a cap 16 for the mechanoprotective frame according to the general inventive concepts. The cap comprises an exterior portion 116a, which faces the exterior of the device and is rounded to prevent potential injury to the subject while the device is implanted. The cap also includes an interior portion 116b, which is a flat portion of the cap that extends from the bottom and has an outer perimeter that is sized and shaped to mate and seal with an inner diameter / perimeter of an opening 120a / b of the mechanoprotective frame. Figure 8B is a bottom view of the cap. Figure 8C is a back view of the cap. Figure 8D is a cross-sectional view of the cap. In embodiments that contain closure caps at the respective longitudinal ends of the frame, said caps have a length suitable for the function of sealing to, e.g., the frame, while having transverse dimensions similar to those of the frame. If deemed necessary or desirable, additional fastening elements may be provided to suitably secure the plunger, and / or cap in place. In certain exemplary embodiments where a plunger is provided, and surgically removed following vascularization, the open end of the frame is thereaftersuitably closed with, e.g., another cap, and the surgical opening is likewise suitably closed. The cap may be constructed from a medical grade metal (stainless steel or titanium), plastic, Teflon®, or medical grade USP Class VI silicone, high consistency silicon rubber (HCR), or liquid silicon rubber (LSR).

[0088] Figure 9A is a top view of an embodiment of a device 100 according to the general inventive concepts. The figure shows the cap 116 removed from the lower opening 120a of the mechanoprotective frame 110. Figure 9B is a perspective view of the device showing the cap positioned / inserted in the opening. Figure 9C is a back view of the device of Figure 9A. Figure 9D is a side cross-sectional view of the device of Figure 9A. Figure 9E is a bottom view of the device of Figure 9 A.

[0089] Figure 10A is a top view of a mechanoprotective frame 110 for a device according to the general inventive concepts. Figure 10B is a perspective view of the mechanoprotective frame. Figure 10C is a back view of the mechanoprotective frame showing an opening 120b. Figure 10D is a front view of the mechanoprotective frame shwonignan opening 120a. Figure 10E is a side cross-sectional view of the mechanoprotective frame. Figure 10F an enhanced view of an embodiment of the mesh wall of the mechanoprotective frame. In certain embodiments, the mesh wall comprises perforations or holes. The perforations in the frame may be, e.g., 100-1000 microns, 200-600 microns, or more preferably 300-500 microns. By way of example, a stainless steel mesh with holes of about 400 microns (diameter) may be provided, but the holes could be slightly smaller or bigger depending on the therapeutic needs among other factors.

[0090] Figure 11 is a perspective view of a plunger 112 which is disposed within the cavity during the vascularization phase and defines a vascularization volume or gap with the frame of about 1-2 mm around the perimeter of the portion of the plunger that is engaged in the frame (114c). The plunger comprises a cap end 114a, which operates in a similar fashion to the cap 116 and an insertion end 114b, which is inserted into the mechanoprotective frame during the vascularization phase. The insertion end of through-hole 113 is shown. The plunger includes a rounded rectangular portion 114c extending between the cap end and the insertion end. As previously mentioned the exterior surface of this portion of the plunger is sized and shaped to mimic the shape of the frame, albeit with reduced dimensions. In this regard, it is preferred that the size of the plunger be limited to allow for 1 to 2 mm of capillary ingrowth all around the plunger, inside the frame. The plunger is preferably a solid component having a shapegenerally corresponding to that of the mechanoprotective frame. The plunger may however have a slightly different shape than the outer mechanoprotective frame to facilitate insertion and removal. For example, the walls of the plunger may be slightly tapered in the insert direction and / or may be grooved or surface treated to facilitate positioning / removal. The plunger may be solid or hollow, although solid (except for the through-hole) is preferred for dimensional stability. The plunger may be made of a medical grade plastic, PTFE, Teflon®, or medical grade USP Class VI silicone, high consistency silicon rubber (HCR), or liquid silicon rubber (LSR). In certain exemplary embodiments, once the vascularization has sufficiently progressed, the plunger is accessed and then slidably removed from within the cavity. The biological material for transplantation, e.g. therapeutic cells / tissue and / or products thereof, is then disposed within the cavity (e.g., via the through-hole) previously occupied by the plunger. In accordance with another embodiment of the invention, the device is implanted already loaded with biological material, e.g. cells / tissue and / or products thereof, and without any plunger structure.

[0091] Figure 12A is a top view of a plunger according to the general inventive concepts, showing the cap end 114a, an insertion end 114b, the insertion end of through-hole 113 rounded rectangular portion 114c extending between the cap end and the insertion end. Figure 12B is a cross-sectional view of the plunger. Figure 13A is a back view of the plunger (i.e., from the perspective of the insertion end 114b). Figure 13B is a front view of the plunger (i.e., from the perspective of the cap end 114a).

[0092] Figure 14 is an image of a device according to the general inventive concepts. The device comprises a mechanoprotective shell and a plunger (shown separated from the frame / shell. In certain embodiments, the loading capacity of the device / plunger is approximately 150 microliters. In certain embodiments, the mechanoprotective shell comprises perforations / holes with a diameter of approximately 400 microns.

[0093] Figure 15A is an image of a device showing the plunger removed from the mechanoprotective frame. The frame is shown from a back perspective with a cap shown positioned in the opening. Figure 15B is an image showing the device assembled with the plunger positioned within the adjacent space.

[0094] Figure 16A is a perspective view of an alternative embodiment of a plunger 122. In certain exemplary embodiments, plunger 122 does not comprise a through-hole. In certainembodiments, the plunger 122 is a two-piece rectangular apparatus, having first end 123 that terminates (in the length direction) in a narrower insertion point, sides in the width direction that are rounded in a fashion similar to the mechanoprotective frame, and a grip portion 124 at the second end. The removeable placeholder / plunger forms a hollow conduit to allow for cell / tissue loading that will occur progressively as the placeholder is withdrawn after vascularization. In this embodiment, the outer portion of the plunger forms a hollow rectangular tube that wraps around the inner placeholder 125. Thus, removal of the inner placeholder 125 provides a hollow inner volume in the device for cell / tissue loading upon removal. Figure 16B is a front view of the plunger shown in 16A. The plunger may be constructed from a medical grade plastic, Teflon®, or medical grade USP Class VI silicone, high consistency silicon rubber (HCR), or liquid silicon rubber (LSR).

[0095] Figure 17A is a top view of the plunger 122 shown in 16A, the first end 123 terminates (in the length direction) in a narrower insertion point. Figure 17B is a cross-sectional view of the plunger shown in 16A where the inner placeholder 125 is shown extending the length of the plunger 122.

[0096] The general inventive concepts also recognize that in addition to providing nutrients, factors, cytokines, drugs, and the like through the cavity during biomaterial implantation, the mechanoprotective frame and / or the plunger (if provided) may be coated with a suitable media, such as a biocompatible polymer impregnated with suitable drug(s) and / or factor(s) to also act as a regulated or unregulated drug delivery system, particularly when the device is first implanted.

[0097] Thus, the general inventive concepts enable immunosuppression-free biologic cures, applicable to a variety of sources of islets from pancreatic, stem-cell derived, or gene edited (hypo-immune) islets, or other therapeutic cell / tissue products by a combination strategy integrating 1) a vascularizing device that allows for blood oxygen / nutrients delivery to therapeutic implanted tissues / cells (VADIT); 2) carrier matrix for prolonged (e.g., >1 week) local delivery of tolerance inducing drugs, cells or other biologies that can be pre-loaded into the MF AT drug delivery system; and 3) a therapeutic cell / tissue product, such as an islet product.

[0098] In certain exemplary embodiments, the general inventive concepts are embodied in a method for creating a vascular bed to define a reservoir for receiving biological material (e.g.,carrier matrix and therapeutic cells) and for facilitating long term survival and function of the biological material within the frame, the method comprises implanting the a device comprising a mechanoprotective frame in the body of the subject with the plunger (when provided) disposed inside or adjacent to the mechanoprotective frame to define a gap or volume for tissue ingrowth. The implant location may be, for example, intraomental (an omental pouch), hepatic, subcutaneous, intraperitoneal, intramuscular, or renal subcapsular. After some time for vascularization in and around the device (e.g., 1-8 weeks), the plunger is accessed and removed and the selected therapeutic cells are deposited in the volume previously occupied by the plunger. In certain exemplary embodiments, the therapeutic cells are implanted via the through- hole while the plunger is withdrawn / removed. In any of the embodiments, the biological material may be implanted in a one-step or a two-step procedure.

[0099] In certain exemplary embodiments, the general inventive concepts are embodied in a method for implanting biological material in a patient in a two-step procedure, comprising: implanting a device for receiving biological material (i.e., a device according to any of the embodiments described herein) at a selected location within the patient, said device including a mechanoprotective frame defining an adjacent, e.g. inner, space; a plunger occupying a part of the adjacent space; allowing tissue ingrowth into the adjacent space; removing the plunger and disposing a biological material comprising, for example, a therapeutic tissue / cell product within the adjacent space vacated by the plunger, and wherein the selected tissue / cell product is provided in a carrier matrix; and delivering at least one of an immunosuppressive and / or growth factor media to said adjacent space. In certain exemplary embodiments, the biological material comprises pancreatic islet cells, SA-FasL, and the carrier matrix, e.g., MF AT. In certain embodiments in which the biological material comprises pancreatic islets for insulin production, the amount of cells generally desired for the treatment of diabetes referred to hereinabove is about 4,000 islets about to 15,000 islets per kilogram of the subject’s weight, including about 4200 islets to about 14,000 islets, including about 4500 islets to about 13,000 islets, including about 5000 islets to about 12,000 islets per kilogram of the subject’s weight.[000100] In certain exemplary embodiments, the general inventive concepts contemplate a method of implanting biological material in a patient via the systems and devices described herein. The method comprises implanting into the patient a device comprising a perforated mechanoprotective frame defining an inner space for receiving biological material, the perforated mechanoprotective frame comprising a first opening and a second opening opposingthe first opening, a plunger positioned in the inner space of the frame and which is removable before deposition of the biological material, an outer surface of the plunger defining a vascularization volume with an inner surface of the perforated mechanoprotective frame, and a cap positioned in a first opening in the perforated mechanoprotective frame opposite to the plunger; allowing tissue ingrowth into and / or around the inner space of the device, accessing the device and removing the plunger element and depositing the biological material into the space vacated by the plunger element, the biological material comprising a therapeutic cell, a carrier matrix, and a tolerance inducing drug, cell, or other biologic; and positioning a cap element in an open end of the device.[000101] The invention may also be embodied in a method for implanting biological material in a patient, wherein prior to disposing the biological material within the space defined by the device, the biological material is encapsulated to provide a means to distribute, give structural integrity, and / or immunoprotect the cells. In certain embodiments, the biological material comprises a therapeutic cell / tissue or product thereof providing therapeutic benefit to the patient when implanted.[000102] The invention may also be embodied in a method for implanting biological material in a patient, wherein the biological material disposed in the space defined by the device comprises, for example, autologous, heterologous, syngeneic, allogeneic, or xenogeneic cells / tissue. The cells may be derived from cadaver tissue or from living tissue. The cells may be of non-mammalian or mammalian origin, non-human origin or human origin, self or nonself. The cells may be pluripotent, multipotent, totipotent, or differentiated embryonic or adult stem cells; primary differentiated cells; or immortalized cells, among other cell types. Stem cells may comprise, e.g., cells derived from cord blood, amniotic fluid, menstrual blood, placenta, Wharton's jelly, cytotropoblasts, and the like. The biological material may also comprise any combination of the above-listed cell types. Biological materials of the invention comprise or consist essentially of the above-listed cell types, or may consist of the above-listed cell types.[000103] To further increase the effectiveness of the treatment, the biological material and / or therapeutic cells may comprise or consist essentially of factor-producing cells that have been genetically manipulated by known techniques to produce one or more therapeutic effects on the patient, such as a secreted therapeutic factor. In the present invention, these may be combined with one or more helper cells or cell types, e.g., Sertoli cells, in order toimmunologically protect the islets from host immune-mediated rejection. In addition, or in the alternative, cells disposed within the device may include cells that produce substances with a different therapeutic activity as in the case of thyroid and parathyroid cells, among others.[000104] Exemplary therapeutic factors which may be delivered by the transplanted cells include, but are not limited to, one or more of: insulin, other hormones or enzyme replacement therapies In some embodiments, the therapeutic factor has insulin-like or insulin-regulatory activity. In certain embodiments, the therapeutic factor is insulin. In certain embodiments, the therapeutic factor is a precursor form of insulin, such as preproinsulin or proinsulin.[000105] In some embodiments, the therapeutic effect comprises regulation of insulin levels in the blood. In certain embodiments, the therapeutic effect comprises regulation of glucose levels in the blood. In other embodiments, the therapeutic effect comprises regulation of levels of one or more other biological response regulators in the blood of the patient.[000106] Besides pancreatic islets, which are considered one preferred therapeutic cell / tissue type for regulating sugar and energy metabolism, and for treating diabetes, the devices of the general inventive concepts and methods involving those devices may also be applied to other tissue and cell therapy model systems. Tissues and cells for implantation may deliver a therapeutic benefit, e.g. by expressing a therapeutic factor in vivo. Many more beneficial cell produced factors or cellular / tissue activities may be imagined. The implanted tissues or cells may express and / or deliver more than one therapeutic factor, or may comprise two or more cell types delivering one or more therapeutic factors. The implanted tissues or cells may also or alternatively express and / or deliver an agonist, analog, derivative, chimera, fusion, or fragment of a therapeutic factor to deliver a therapeutic effect.[000107] The implanted tissues or cells may also or alternatively deliver a therapeutic effect without secreting a diffusible factor, e.g. by providing an enzymatic activity that, for example, converts a substrate into a product having a beneficial effect, and / or metabolizing, sequestering, or absorbing a detrimental substance. The implanted tissues or cells may deliver a therapeutic effect through a biological material-linked factor, such as a cell surface-linked factor.[000108] The tissues or cells may naturally deliver a therapeutic effect, without genetic modifications, or may be genetically engineered to do so. For example, the biological material of the invention may comprise therapeutic cells transfected with expression vectors that express one or more therapeutic and / or helper cell factors. In another embodiment, the biologicalmaterial of the invention may consist essentially of cells transfected with expression vectors that express one or more therapeutic and / or helper cell factors. In another embodiment, the biological material of the invention may consist of cells transfected with expression vectors that express one or more therapeutic and / or helper cell factors. Such expression may be in a constitutive or in a regulated manner, e.g., in response to biological modulators in the bloodstream or tissues to which the hybrid device is exposed.[000109] It is a further object of the invention to provide a receptacle for implanted biological material that favors cellular survival by providing mechanical support while (a) maximizing exposure of the transplant to new capillaries growing within and / or around the device (for example, by delivery of VEGF or VEGF pathway agonists or the use of degradable, angiogenic materials); and (b) locally delivering substances that can promote not only growth of new capillaries but also protect / enhance the implanted biological material, e.g. cells / tissues and / or products thereof (such as, e.g., anti-inflammatory, antiapoptotic products and / or growth factors such as corticosteroids (e.g., prednisolone, dexamethasone, loteprednol etabonate, flucinolone acetonide, etc.), IGF-I, IGF-II, HGF, GLP-1, Exendin-4, INGAP, lysophylline, pentoxyfilline; COX-2 inhibitors; interleukin-1 receptor antagonist peptide (IRAP), interleukin- 10 (IL-10), alpha 1 -antitrypsin (AAT), TGF-beta; antibodies to IL-1, interferon-gamma, and TNF-alpha; anti-tissue factor, complement inhibitors, oxygen generating, releasing (such as encapsulated peroxides), or transport-enhancing (such as perfluorocarbon PFC) products; as well as endothelial progenitor cells, stem cells, regulatory T cells Treg, or any others known to those skilled in the art, which may optionally or additionally be encapsulated.[000110] In some embodiments, the device of the invention is packaged in a sterile packaging optionally including a label and / or instructions for use of the device. Preferably, the sterile, prepackaged device is ready for use according to one or more of the methods of the invention. Devices of the invention may but need not necessarily be associated with a biomaterial when the sterilization step is performed. As the skilled artisan will readily appreciate, when one or more biomaterials are associated with the device before it is sterilized, the sterilization method is preferably selected to preserve the activity and / or viability of the biomaterial. Alternatively, devices of the invention may be sterilized before they are associated with a biomaterial according to the invention.[000111] The following examples illustrate features and / or advantages of the compositions, systems, and methods according to the general inventive concepts. The examples are givensolely for the purpose of illustration and are not to be construed as limitations of the general inventive concepts, as many variations thereof are possible without departing from the spirit and scope of the general inventive concepts.[000112]In certain exemplary embodiments, the general inventive concepts comprise the following procedure(s): the MF AT will undergo two washes with PBS via centrifugation (200 gxlO); subsequently, 20 pL of MF AT will be mixed with 2, 4, or 10 pg of SA-FasL solution for small subjects (e.g., rodent). For larger subjects (e.g., non-human primates etc.) the MF AT will be mixed with 200-300 micrograms od SA-FasL. For human subjects, the MFAT will be mixed with 3-8 milligrams of SA-FasL. The resulting mixture will be agitated gently for 30 min. In certain exemplary embodiments, the MFAT and islet are provided in a MFAT to IEQ (islet equivalents) ratio of about 3 : 1 to about 1 : 1, including a ratio of 2: 1.[000113]Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the embodiments belong. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the various embodiments, the preferred methods and materials are described herein. All references cited herein, including published or corresponding U.S. or foreign patent applications, issued U.S. or foreign patents, or any other references, are each incorporated by reference in their entireties, including all data, tables, figures, and text presented in the cited references. In the drawings, the thickness of the lines, layers, and regions may be exaggerated for clarity. It is to be noted that like numbers found throughout the figures denote like elements. The terms “system” and “inventive system” may be used interchangeably herein.[000114] The systems and corresponding methods of the present disclosure can comprise, consist of, or consist essentially of the essential elements and limitations of the disclosure as described herein, as well as any additional or optional ingredients, components, or limitations described herein or otherwise useful in transplantation related applications.[000115]The systems and corresponding methods of the present disclosure may also be substantially free of any optional or selected element or feature described herein, provided that the remaining composition still contains all of the required elements or features as described herein. In this context, and unless otherwise specified, the term “substantially free” means thatthe selected composition contains less than a functional amount of the optional ingredient, typically less than 0.1% by weight, and also including zero percent by weight of such optional or selected essential ingredient.[000116] While the invention has been illustrated and described in detail in the drawings and foregoing description, the same is to be considered as illustrative and not restrictive in character. It should be understood that only the exemplary embodiments have been shown and described and that all changes and modifications that come within the spirit of the invention are desired to be protected.

Claims

Claims:

1. A system for the implantation of therapeutic tissues, the system comprising: a device including a perforated mechanoprotective frame defining an inner space for receiving biological material, the perforated mechanoprotective frame comprising a first opening and a second opening opposing the first opening, a plunger positioned in the inner space of the frame and which is removable before deposition of the biological material, an outer surface of the plunger defining a vascularization volume with an inner surface of the perforated mechanoprotective frame, and a cap positioned in a first opening in the perforated mechanoprotective frame opposite to the plunger; and wherein the biological material comprises a therapeutic cell, a carrier matrix, and a tolerance inducing drug, cell, or other biologic.

2. The system of claim 1, wherein the device has a thickness of about 0.4 cm to 1 cm, preferably about 0.5 cm to about 0.7 cm, and more preferably about 0.6 cm.

3. The system of claim 1, wherein the device has a capacity of about 4 ml to about 15 ml for biological material.

4. The system of claim 1, wherein the vascularization volume has a depth of about 0.5 mm to about 1.5 mm around the plunger.

5. The system of claim 1, wherein the plunger comprises a through-hole connecting an exterior of the device to the inner space.

6. The system of claim 1, wherein the plunger is comprised of medical grade plastic, PTFE, Teflon®, or medical grade USP Class VI silicone, high consistency silicon rubber (HCR), or liquid silicon rubber (LSR).

7. The system of claim 1, wherein the mechanoprotective frame is comprised of stainless steel, titanium, or biocompatible plastic material.

8. The system of claim 1, wherein the mechanoprotective frame comprises perforations having a diameter of about 100 microns to about 1000 microns, preferably about 200 microns to about 800 microns, more preferably about 300 microns to about 600 microns, and most preferably about 400 microns.

9. The system of claim 1, wherein the device further comprises a tether for accessing the device after implantation.

10. The system of claim 1, wherein the therapeutic cell is selected from pancreatic islets, adipose tissue products, mesenchymal stromal cells, and subcellular products such as exosomes and extracellular vesicles (EVs).

11. The system of claim 1, wherein the carrier matrix is MF AT, and the tolerance inducing drug, cell, or other biologic is SA-FASL.

12. The system of claim 11, wherein the therapeutic cell is a pancreatic islet and MF AT and pancreatic islet are provided in a MF AT to islet equivalent (IEQ) ratio of about 3 : 1 to about 1 : 1.

13. The system of claim 1, wherein the therapeutic cell is provided in an amount of 4,000 to 15,000 islets per kilogram of a subject’s weight.

14. A method of implanting biological material in a patient, comprising the steps of: implanting into the patient a device comprising: a perforated mechanoprotective frame defining an inner space for receiving biological material, the perforated mechanoprotective frame comprising a first opening and a second opening opposing the first opening, a plunger positioned in the inner space of the frame and which is removable before deposition of the biological material, an outer surface of the plunger defining a vascularization volume with an inner surface of the perforated mechanoprotective frame, and a cap positioned in a first opening in the perforated mechanoprotective frame opposite to the plunger; and allowing tissue ingrowth into and / or around the inner space of the device,accessing the device and removing the plunger element and depositing the biological material into the space vacated by the plunger element, the biological material comprising a therapeutic cell, a carrier matrix, and a tolerance inducing drug, cell, or other biologic; and positioning a second cap in an open end of the device.

15. The method of claim 14, wherein the therapeutic cell is selected from pancreatic islets, adipose tissue products, mesenchymal stromal cells, and subcellular products such as exosomes and extracellular vesicles (EVs).

16. The method of claim 14, wherein the carrier matrix is MF AT.

17. The method of claim 14, wherein the tolerance inducing drug, cell, or other biologic is selected from rapamycin or anti-CD40L antibodies, PD-L1, SA FasL or other FasL soluble or oligomeric formulations.

18. The method of claim 14, wherein the therapeutic cell is pancreatic islets, the carrier matrix is MF AT, and the tolerance inducing drug, cell, or other biologic is SA-FASL.

19. The method of claim 18, wherein the MF AT and pancreatic islet are provided in a MF AT to islet equivalent (IEQ) ratio of about 3 : 1 to about 1 : 1.

20. The method of claim 14, wherein the therapeutic cell is provided in an amount of 4,000 to 15,000 islets per kilogram of a subject’s weight.

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