Bioartificial Endocrine Pancreas Research Articles

47: Bioartificial Endocrine Pancreas Generated from Decellularized Human Placenta for Type-1 Diabetes Treatment

Background Development of vascularized biological scaffolds providing mechanical protection for islets is a challenging objective of modern regenerative medicine. The aim of our study was to develop vascularized endocrine pancreas using decellularized placental cotyledon. Methods/ Materials Human placentas were decellularized by perfusion, using 0.5% sodium dodecyl sulfate. Decellularization was assessed by histological analyses, scanning electron microscopy (SEM) and residual DNA quantification. Glucosaminoglycans (GAG) and hydroxyproline were analyzed. Placental cotyledons were recellularized by a intravascular perfusion of human umbilical vein endothelial cells (HUVECs) during the 24h, followed by intraparenchimal injection of INS-1E cells. Recellularization was assessed by hystological and immunohistochemical methods, and endocrine function was confirmed by glucose stimulated insulin secretion tests performed at different time points. Results Histological staining and SEM showed complete decellularization and well preserved ECM structure. This was confirmed by the absence of residual DNA. Immunostainings of recellularized cotyedons showed insulin expressing islet like structures with intense vascularization confirmed by CD31 staining. Adequate insulin secretion in response to high glucose stimulation was also observed, confirming the functional activity of INS-1E cells. Conclusion These data demonstrate that acellular placental cotyledons seeded with endocrine pancreatic tissue and endothelial cells could be used for functional pancreas bioengineering.

The Human Pancreas as a Source of Protolerogenic Extracellular Matrix Scaffold for a New-generation Bioartificial Endocrine Pancreas.

Our study aims at producing acellular extracellular matrix scaffolds from the human pancreas (hpaECMs) as a first critical step toward the production of a new-generation, fully human-derived bioartificial endocrine pancreas. In this bioartificial endocrine pancreas, the hardware will be represented by hpaECMs, whereas the software will consist in the cellular compartment generated from patient's own cells. Extracellular matrix (ECM)-based scaffolds obtained through the decellularization of native organs have become the favored platform in the field of complex organ bioengineering. However, the paradigm is now switching from the porcine to the human model. To achieve our goal, human pancreata were decellularized with Triton-based solution and thoroughly characterized. Primary endpoints were complete cell and DNA clearance, preservation of ECM components, growth factors and stiffness, ability to induce angiogenesis, conservation of the framework of the innate vasculature, and immunogenicity. Secondary endpoint was hpaECMs' ability to sustain growth and function of human islet and human primary pancreatic endothelial cells. Results show that hpaECMs can be successfully and consistently produced from human pancreata and maintain their innate molecular and spatial framework and stiffness, and vital growth factors. Importantly, hpaECMs inhibit human naïve CD4 T-cell expansion in response to polyclonal stimuli by inducing their apoptosis and promoting their conversion into regulatory T cells. hpaECMs are cytocompatible and supportive of representative pancreatic cell types. We, therefore, conclude that hpaECMs has the potential to become an ideal platform for investigations aiming at the manufacturing of a regenerative medicine-inspired bioartificial endocrine pancreas.

Capturing Islet Stem Cells for a Bio-Artificial Pancreas.

Encapsulation cell therapy, allows for embryonic pancreatic islet cells to differentiate into functional alpha, beta, and delta cells, mimicking the normal composition of the islets (“bio-artificial endocrine pancreas”) would be ideal for the treatment of diabetes. The use of immunosuppressive medications in allo-islet or pancreatic transplant has been associated with increased infections and malignancy. The goal of encapsulation technology is to safely protect the transplanted islets from both allo-rejection and autoimmunity without immunosuppression. The device prevents rejection by encapsulating the progenitor cells in a semi-permeable membrane that allows smaller molecules, such as oxygen, insulin, glucose and albumin to freely pass through the membrane. At the same time, the membrane prevents larger molecules, such as immune cells and antibodies, from entering the capsule, thereby preventing rejection of the differentiating and developing islet cells. The advent of encapsulation products that could cure Type 1 diabetes raises many questions. A few of those questions on the encapsulation cell therapy are reviewed in this article.

Design for a Lithographically Patterned Bioartificial Endocrine Pancreas

Cell encapsulation provides a means to transplant therapeutic cells for a variety of diseases including diabetes. However, due to the large numbers of cells, approximately on the order of a billion, that need to be transplanted for human diabetes therapy, adequate mass transport of nutrients such as oxygen presents a major challenge. Proof-of-concept for the design of a bioartificial endocrine pancreas (BAEP) that is optimized to minimize hypoxia in a scalable and precise architecture is demonstrated using a combination of simulations and experiments. The BAEP is composed of an array of porous, lithographically patterned polyhedral capsules arrayed on a rolled-up alginate sheet. All the important structural variables such as the capsule dimensions, pore characteristics, and spacing can be precisely engineered and tuned. Further, all cells are encapsulated within a single device with a volume not much greater than the total volume of the encapsulated cells, and no cell within the device is located more than 200 μm from the surrounding medium that facilitates efficient mass transport with the surroundings. Compared with gel-based encapsulation methods, our approach offers unprecedented precision and tunability of structural parameters as well as the volume of the encapsulated cells and consequently the amount of secreted insulin. Our work highlights the utility of lithography and self-assembly in the fabrication of micro- and nanostructured three-dimensional structures that simulate the function of natural endocrine organs.

Improvement of Hyperglycemia and Sexual Dysfunction in Diabetic Female Rats by an Artificial Endocrine Pancreas Developed from Mouse .BETA. Cells

We investigated the effects of a bioartificial endocrine pancreas (Bio-AEP) produced by mouse beta cells on sexual dysfunction of streptozotocin (STZ)-induced diabetic female rats. Female rats were administered STZ (60 mg/kg BW, i.v.) at the age of 10 weeks and transplanted with a Bio-AEP including mouse beta cells at the age of 14 weeks (STZ+Bio-AEP group). Lordosis and proceptive sexual behavior of female rats were observed. The results showed that after the Bio-AEP transplant blood glucose recovered from 380-450 mg/dl induced by streptozotocin to 140-230 mg/dl and suppressed lordosis and proceptive behavior also recovered. These results suggest that it is possible to reverse sexual dysfunction by continuous administration of mouse insulin.

Unusual Permeability of Porcine Endogenous Retrovirus Subgroup A Through Membrane Filters

In xenotransplantation from pigs to humans, a bio-artificial endocrine pancreas (Bio-AEP), in which pancreatic endocrine cells are encapsulated within a semipermeable membrane of 100 nm pore size, has been developed. We evaluated the permeability of porcine endogenous retroviruses (PERVs) through membrane filters using a pseudotype virus (LacZ(PERV-A)) containing a viral core derived from murine leukemia virus and an envelope (Env) from PERV subgroup A. Contrary to our expectations, LacZ(PERV-A) lost its infectivity by filtration through a 200 nm membrane filter. This unusual phenotype was not observed in pseudotype viruses harboring Envs from other gammaretroviruses. The infectivity of LacZ(PERV-A) was significantly decreased by repeated freeze/thaw treatment, indicating that LacZ(PERV-A) was physically labile. In addition, LacZ(PERV-A) may be agglutinated because copy numbers of viral RNA after filtration were significantly reduced by filtration through the 200 nm membrane. This phenotype is advantageous to develop a safe Bio-AEP blocking PERV infection.

Diabetes mellitus: rational basis, clinical approach and future therapy

Clinical islet transplantation has recently received a strong impulse from the results obtained with the introduction of the Edomonton group. However, islet transplantation is at present a minimally invasive procedure and offers for the future the unique possibility of being performed under donor-specific tolerant conditions because islets may potentially be engineered in vitro. In addition, various approaches such as in vitro islet expansion, or xenogenic islets could make the availability of donor tissues unlimited. Recent advances in tissue engineering (technology) and cell biology may allow for the development of novel strategies for the treatment and cure of type I diabetes. In particular, it is now possible to envisage restoration of insulin secretion by cell-replacement therapy. And it will be necessary to ensure that implanted β-cells are protected in some way from recognition by the immune system (a bio-artificial endocrine pancreas).

Preclinical development of the Islet Sheet.

The Islet Sheet is a thin planar bioartificial endocrine pancreas fabricated by gelling highly purified alginate and islets of Langerhans. Acellular alginate layers form a uniform immunoprotective barrier to host rejection of the encapsulated cells, with the tissue nourished by passive diffusion from adjacent host tissue. The overall thickness of the Islet Sheet, 250 microm, is chosen to maximize nutrient diffusion. In this paper we describe the early development of the Islet Sheet, including purification and fractionation of the alginates used, difficulties in maintaining sheet planarity, and preliminary metabolic studies in pancreatectomized dogs. In a key experiment, approximately 75,000 allogeneic islet equivalents in six Islet Sheets were sutured to the omentum of a 7-kg female beagle dog at the time of pancreatectomy. Fasting euglycemia was maintained for 84 days. Fed blood sugars were usually below 150 mg/dL. A single injection of 2 U insulin was administered on day 9, and antibiotics were administered for two weeks. No other drugs were used. IVGTT post implant was not normal, but seemed to improve between 30 and 60 days. Upon omentectomy and sheet removal the metabolic parameters deteriorated to a frankly diabetic state within seven days. The sheets did not remain flat, but fragments were recovered within hard, mostly acellular capsules. Dithizone staining showed islets within alginate sheets recovered from the interior of these capsules, suggesting that allogeneic islet tissue survived 84 days and was responsible for maintaining fasting euglycemia.

Bioartificial pancreas: materials, devices, function, and limitations.

The term "bioartificial endocrine pancreas" (BEP) was introduced by Anthony Sun in 1980. It was in 1968, however, that Thomas Chang proposed the use of microencapsulated islets as artificial beta-cells. By applying a semipermeable membrane on the top of microcapsules, a system can be produced that is impermeable to viable islet cells and large effector molecules of the immune system, thus providing a protection for transplanted islets against rejection. Since then, the term BEP has not often appeared in papers. Instead, the term "bioartificial pancreas" (BAP) has gained widespread use. In a broader sense, BAP would include an application of suitable endocrine cells and protective polymeric vehicles, but not necessarily providing a filtration barrier of precisely defined properties (e.g., cells injected into a gel of hyaluronate).

Biocompatibility of two different types of extracellular matrix in bioartificial endocrine pancreas in dogs: A preliminary report

The biocompatibility of synthetic extracellular matrix (ECM) in the diffusion chamber of a bioartificial endocrine pancreas (Bio-AEP) was investigated in normal beagle dogs. Two different types of ECM were used in this study, type I collagen treated with pepsin to ensure lower antigenicity (type I LA) and reconstituted basement membrane matrix (Matrigel derived from Englbreth-Holm-Swarm mouse sarcoma. Bio-AEPs with type I LA or Matrigel without pancreatic endocrine cells were implanted into the abdominal cavities of beagle dogs (n=4). At 1 month after implantation, laparoscopy revealed no abnormalities in the Bio-AEPs with type I LA, whereas it revealed encapsulation of one of the Bio-AEPs with Matrigel with fibrous tissue, and in that case the Bio-AEP was removed. At 3 months after implantation, the remaining Bio-AEP with Matrigel was completely destroyed, whereas the Bio-AEPs with type I LA showed no abnormal findings. Microscopically, an invasion of red blood cells and macrophages into the chamber was observed in the Bio-AEPs with Matrigel. A proliferation of fibroblasts around the Matrigel was observed in one case. In contrast, light microscopic findings revealed no abnormal findings in the Bio-AEP with type I LA. These results suggest that the cell growth and differentiation factors and adhesion molecules in Matrigel may play an important role in fibrous encapsulation and may be incompatible with dogs, and that type I LA is more suitable for Bio-AEP.

Effect of the Extracellular Matrix on Pancreatic Endocrine Cell Function and its Biocompatibility in Dogs

The effect of the synthetic extracellular matrix (ECM) in a diffusion chamber for a bioartificial endocrine pancreas (Bio-AEP) on pancreatic endocrine cells in vitro and its biocompatibility in dogs were investigated. Two different types of ECM were used: type I collagen treated with low antigen (type I LA), and reconstituted basement membrane matrix (Matrigel) derived from Englbreth-Holm-Swarm (EHS) mouse sarcoma. Matrigel contains growth and differentiation factors and cell adhesion molecules such as laminin, heparan sulfate proteoglycan, and entactin. Purified porcine pancreatic endocrine (PE) cells were suspended in type I LA or Matrigel and then placed into a 12-well culture plate (4 x 10(7) cells/ml gel/well). The insulin accumulation from PE cells in Matrigel was significantly greater than that in type I LA (9.3 +/- 3.6 mU/well vs. 2.3 +/- 1.3 mU/well). When Bio-AEP with Matrigel and PE cells was implanted into the abdominal cavity of a pancreatectomized diabetic dog, the exogenous insulin requirement for maintaining normoglycemia was reduced for the first 4 weeks. However, after 6 weeks of implantation, fasting blood glucose levels suddenly increased. Laparotomy revealed encapsulated Bio-AEP with thick fibrous tissue. Following removal of the Bio-AEP from the abdominal cavity, another Bio-AEP containing type I LA and PE cells was implanted into the same dog. The exogenous insulin requirement was gradually decreased to almost half that of preimplantation levels. Bio-AEPs containing type I LA or Matrigel, but not PE cells, were implanted into the abdominal cavities of four healthy dogs. After 4 weeks of implantation, the Bio-AEP with Matrigel was encapsulated with fibrous tissue similar to that in the diabetic dog, but the Bio-AEP with type I LA was not. These results indicate that Matrigel may be incompatible with dogs and that the type I LA is more suitable for Bio-AEP.

Strategies for immunoisolation in islet transplantation: challenges for the twenty-first century.

A number of investigations have pursued the feasibility of a bioartificial endocrine pancreas (Bio-AEP) in which pancreatic endocrine cells (PE-cells) are encapsulated within a semipermeable membrane to protect them from immunological rejection. The goal of PE-cell transplantation in human diabetes is to maintain the PE-cell graft in the recipient without the use of immunosuppression. One approach is to encapsulate the donor PE-cells in a semipermeable membrane. A type of diffusion chamber for a Bio-AEP is described. The system is based on a chamber designed to improve the diffusion of insulin secretion in response to glucose transfer across a membrane. An implantable diffusion chamber for the Bio-AEP was constructed by placing PE-cells in agarose, containing nicotinamide and a sugar-chain coated polymer, in a ring holder which was sandwiched between two nucleopore membranes (pore size 0.1 microm), for transplantation into diabetic animals. This compatible diffusion chamber prevented rejection, and maintained normoglycemia in diabetic rats when implanted intraperitoneally.

Bioartificial endocrine pancreas (Bio-AEP) for treatment of diabetes: effect of implantation of Bio-AEP on the pancreas.

Recently, we described a diffusion chamber for a bioartificial endocrine pancreas (Bio-AEP). Pancreatic islet cells in the Bio-AEP device were isolated from the immune system of the host by an artificial barrier, while nutrients, electrolytes, oxygen, and bioactive secretory products were exchanged across this barrier. This experiment was designed to evaluate whether the diffusion chamber could be useful as a Bio-AEP in the treatment of diabetes. Six streptozotocin (STZ)-induced diabetic rats each received a diffusion chamber containing 8 x 10(6) MIN6 cells as a xenograft Bio-AEP. In the STZ diabetic rats with Bio-AEPs, a return to normoglycemia was observed up to 30 weeks after implantation, without the use of any immunosuppressant. A gradual increase in the body weight of the rats was also observed. In three STZ diabetic rats, diffusion chambers without MIN6 cells were implanted as a sham operation. The fasting blood glucose levels in these three rats remained higher than 600 mg/dl, after implantation, and they lost weight. Thirty-five weeks after implantation, the pancreata were removed from the rats that underwent xenoimplantation, those that had the sham operation, and the normal control rats. In the sham-operated animals, the exocrine tissues of the pancreata were vacuolated and pancreatic B cells were not seen in the islets. In contrast, in the pancreata from the xenoimplantation, the exocrine tissues were normal, and a few pancreatic B cells were seen in the islets. These results indicated that xenoimplantation using the Bio-AEP might retard the progress of diabetes.

In vitro and in vivo performance of porcine islets encapsulated in interfacially photopolymerized poly(ethylene glycol) diacrylate membranes.

The usefulness of interfacial photopolymerization of poly(ethylene glycol) (PEG) diacrylate at a variety of concentrations and molecular weights to form hydrogel membranes for encapsulating porcine islets of Langerhans was investigated. The results from this study show in vitro and in vivo function of PEG-encapsulated porcine islets and the ability of PEG membranes to prevent immune rejection in a discordant xenograft model. Encapsulated islets demonstrated an average viability of 85% during the first week after encapsulation, slightly but significantly lower than unencapsulated controls. Encapsulated porcine islets were shown to be glucose responsive using static glucose stimulation and perifusion assays. Higher rates of insulin release were observed for porcine islets encapsulated in lower concentrations of PEG diacrylate (10-13%), not significantly reduced relative to unencapsulated controls, than were observed in islets encapsulated in higher concentrations (25%) of PEG diacrylate. Perifusion results showed biphasic insulin release from encapsulated islets in response to glucose stimulation. Streptozotocin-induced diabetic athymic mice maintained normoglycemia for up to 110 days after the implantation of 5,000-8,000 encapsulated porcine islet equivalents into the peritoneal cavity. Normoglycemia was also confirmed in these animals using glucose tolerance tests. PEG diacrylate-encapsulated porcine islets were shown to be viable and contain insulin after 30 days in the peritoneal cavity of Sprague-Dawley rats, a discordant xenograft model. From these studies, we conclude that PEG diacrylate encapsulation of porcine islets by interfacial photopolymerization shows promise for use as a method of xenoprotection toward a bioartificial endocrine pancreas.

Development of a bioartificial pancreas: I. Long-term propagation and basal and induced secretion from entrapped ?TC3 cell cultures

Bioartificial pancreatic constructs based on immunoisolated, insulin-secreting cells have the potential for providing effective, long-term treatment of type I (insulin-dependent) diabetes. Use of insulinoma cells, which can be amplified in culture, relaxes the tissue availability limitation that exists with normal pancreatic islet transplantations. We have adopted mouse insulinoma betaTC3 cells entrapped in calcium alginate/poly-L-lysine/alginate (APA) beads as our model system for a bioartificial pancreas, and we have characterized the effects of long-term propagation and of glucose concentration step changes on the bioenergetic status and on the metabolic and secretory activities of the entrapped cells. Cell bioenergetics were evaluated nonivasively by phosphorus-31 nuclear magnetic resonance ((31)P NMR) spectroscopy, and metabolic and secretory parameters by assaying cell culture medium. Data indicate that net cell growth occurred between days 3 and 10 of the experiment, resulting in an approximate doubling of the overall metabolic and secretory rates and of the intracellular metabolite levels. Concurrently, a reorganization of cell distribution within the beads was observed. Following this growth period, the measured metabolic and secretory parameters remained constant with time. During glucose step changes in the perfusion medium from a high concentration of 12 to 15 mM to 0 mM for 4.5 h to the same high glucose concentration, the oxygen consumption rate was not affected, whereas insulin secretion was always glucose-responsive. Intracellular nucleotide triphosphates did not change during 0 mM glucose episodes performed early in culture history, but they declined by 20% during episodes performed later in the experiment. It is concluded that the system of APA-entrapped betaTC3 cells exhibits several of the desirable characteristics of a bioartificial pancreas device, and that a correlation between ATP and the rate of insulin secretion from betaTC3 cells exists for only a domain of culture conditions. These findings have significant implications in tissue engineering a long-term functional bioartificial endocrine pancreas, in developing noninvasive methods for assessing construct function postimplantation, and in the biochemical processes associated with insulin secretion.

Development of a bioartificial pancreas: I. Long‐term propagation and basal and induced secretion from entrapped βTC3 cell cultures

Bioartificial pancreatic constructs based on immunoisolated, insulin-secreting cells have the potential for providing effective, long-term treatment of type I (insulin-dependent) diabetes. Use of insulinoma cells, which can be amplified in culture, relaxes the tissue availability limitation that exists with normal pancreatic islet transplantations. We have adopted mouse insulinoma βTC3 cells entrapped in calcium alginate/poly-L-lysine/alginate (APA) beads as our model system for a bioartificial pancreas, and we have characterized the effects of long-term propagation and of glucose concentration step changes on the bioenergetic status and on the metabolic and secretory activities of the entrapped cells. Cell bioenergetics were evaluated nonivasively by phosphorus-31 nuclear magnetic resonance (31P NMR) spectroscopy, and metabolic and secretory parameters by assaying cell culture medium. Data indicate that net cell growth occurred between days 3 and 10 of the experiment, resulting in an approximate doubling of the overall metabolic and secretory rates and of the intracellular metabolite levels. Concurrently, a reorganization of cell distribution within the beads was observed. Following this growth period, the measured metabolic and secretory parameters remained constant with time. During glucose step changes in the perfusion medium from a high concentration of 12 to 15 mM to 0 mM for 4.5 h to the same high glucose concentration, the oxygen consumption rate was not affected, whereas insulin secretion was always glucose-responsive. Intracellular nucleotide triphosphates did not change during 0 mM glucose episodes performed early in culture history, but they declined by 20% during episodes performed later in the experiment. It is concluded that the system of APA-entrapped βTC3 cells exhibits several of the desirable characteristics of a bioartificial pancreas device, and that a correlation between ATP and the rate of insulin secretion from βTC3 cells exists for only a domain of culture conditions. These findings have significant implications in tissue engineering a long-term functional bioartificial endocrine pancreas, in developing noninvasive methods for assessing construct function postimplantation, and in the biochemical processes associated with insulin secretion. © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 66: 219–230, 1999.

Membrane immunoisolation of a diffusion chamber for bioartificial pancreas.

Immunoisolation is a potentially important approach to transplanting islets without any immunosuppressive therapy. The concept of immunoisolation is outlined in systems in which the transplanted tissue is separated from the immune system of the host by an artificial barrier. We previously described a diffusion chamber as a bioartificial endocrine pancreas (Bio-AEP), which was constructed by placing pancreatic endocrine cells, trapped in a mixed matrix, in the center of a ring holder sandwiched between nucleopore membranes, which were shielded by silicone. This experiment was designed to evaluate a suitable pore size for the nucleopore membrane to ensure immunoisolation during xenoimplantation of the Bio-AEP in vitro and in vivo. A nucleopore membrane of pore size 0.1 microm or 0.2 microm was employed as the semipermeable membrane which provided a mechanical barrier between the endocrine pancreas graft and the host immune system. The protective effect of the Bio-AEP from humoral immunity was determined in vitro, using sensitized sheep erythrocytes (EAs). A complement protein did not destroy the cell membranes of the EAs in the diffusion chamber containing the mixed matrix with the nucleopore membrane of 0.1 microm pore size. In an in vivo experiment, 6 streptozotocin (STZ) induced diabetic rats were implanted with Bio-AEPs constructed with nucleopore membranes of pore size 0.1 microm and containing MIN6 cells in the mixed matrix. In the STZ diabetic rats with Bio-AEPs, a return to normoglycemia was observed up to 50 weeks after implantation without the use of any immunosuppressant. Also, the body weights of the rats gradually increased. During the observation, when the Bio-AEPs were removed from the STZ diabetic rats, the blood glucose immediately returned to preimplantation levels, and the body weights of the rats also decreased. The membranes of the Bio-AEPs removed from the STZ diabetic rats showed a very thin layer of fibroblastic cells on the outer surfaces. The results indicated that the Bio-AEP, in which pancreatic endocrine cells were trapped in a mixed matrix and with a 0.1 microm pore size membrane, should be useful for xenoimplantation into diabetic animals and may open a new field in the therapy of human diabetics.

Complement Permeability in Bio-Artificial Endocrine Pancreas Using a Diffusion Chamber

Immunoisolation is a potentially important approach to transplanting islets without any immunosuppressive therapy. This study was designed to investigate whether complement penetrates into the diffusion chamber that we developed for a bioartificial endocrine pancreas (Bio-AEP). The results showed that within 12–24 h, when a Nuclepore membrane (Nuclepore Corp., California) was the sole barrier, complement permeated 100%. However, the Nuclepore membrane used together with a mixed matrix prevented complement penetration. Also, the complement become inactivated during penetration through both the mixed matrix and the Nuclepore membrane with a pore size of 0.1 μm. In conclusion, immunoisolation of cells by a mixed matrix and a Nuclepore membrane of 0.1 μm pore size is a particularly effective method for transplanting xenogeneic cells. Thus, the clinical application of our Bio-AEP shows promise for long-term xenotransplantation without an immunosuppressant.

Complement permeability in bio-artificial endocrine pancreas using a diffusion chamber.

Immunoisolation is a potentially important approach to transplanting islets without any immunosuppressive therapy. This study was designed to investigate whether complement penetrates into the diffusion chamber that we developed for a bioartificial endocrine pancreas (Bio-AEP). The results showed that within 12-24 h, when a Nuclepore membrane (Nuclepore Corp., California) was the sole barrier, complement permeated 100%. However, the Nuclepore membrane used together with a mixed matrix prevented complement penetration. Also, the complement become inactivated during penetration through both the mixed matrix and the Nuclepore membrane with a pore size of 0.1 microm. In conclusion, immunoisolation of cells by a mixed matrix and a Nuclepore membrane of 0.1 microm pore size is a particularly effective method for transplanting xenogeneic cells. Thus, the clinical application of our Bio-AEP shows promise for long-term xenotransplantation without an immunosuppressant.

A Bio-Artificial Endocrine Pancreas for the Treatment of Diabetes

A Bio-Artificial Endocrine Pancreas for the Treatment of Diabetes