Entrapment of Cultured Pancreas Islets in Three-Dimensional Collagen Matrices

In vitro culture of islets of Langerhans decreases their immunogenecity, presumably by eliminating passenger leukocytes and other Ia+ presenting cells within the islets. Islets cultivated in petri dishes either at 37°C or at 25°C gradually disintegrate during culture in a time-dependent manner which is related to the free-floating condition of the islets. Also, a fraction of the islets disperse as single cells and beta-cell aggregates or adhere to the bottom of the culture dishes. Thus, the retrieval rate of transplantable islets is dampened due to their disintegration and spontaneous dispersion in conventional petri dish cultures. Entrapment of freshly harvested islets of Langerhans in a three-dimensional collagen matrix was studied as an alternative method for islet cultivation. The contraction of collagen fibrils during in vitro culture counteracts the dispersion of islets and helps in maintaining their integrity while in culture. It was observed that the entrapped islets maintain satisfactory morphology, viability, and capability of glucose-dependent insulin secretion for over 2 wk. The oxygen consumption rate and glucose metabolism of these islets was not deranged when entrapped in collagen. Also, the retrieval of islets is easier and more efficient than that observed in conventional culture systems. Our results indicate that culture of islets in three-dimensional collagen gels can potentially develop into an ideal system applicable to clinical transplantation of cultured islets or beta-cell aggregates.

[1]  M. Field,et al.  Autotransplantation of dispersed pancreatic islet tissue combined with total or near-total pancreatectomy for treatment of chronic pancreatitis. , 1991, Surgery.

[2]  R. Nakhleh,et al.  Pancreas Transplant Pathology: A Morphologic, Immunohistochemical, and Electron Microscopic Comparison of Allogeneic Grafts with Rejection, Syngeneic Grafts, and Chronic Pancreatitis , 1991, The American journal of surgical pathology.

[3]  G. Charriere,et al.  Tissue reaction to subcutaneous implantation of a collagen sponge. A histological, ultrastructural, and immunological study. , 1990, Journal of biomedical materials research.

[4]  A. E. Dolby,et al.  Development and testing of a human collagen graft material. , 1990, Journal of biomedical materials research.

[5]  P H Robinson,et al.  Two-ply biodegradable nerve guide: basic aspects of design, construction and biological performance. , 1990, Biomaterials.

[6]  J. Markmann,et al.  THE EFFECT OF ISLET CELL CULTURE IN VITRO AT 24°C ON GRAFT SURVIVAL AND MHC ANTIGEN EXPRESSION , 1990 .

[7]  H. Iwata,et al.  Evaluation of Microencapsulated Islets in Agarose Gel as Bioartificial Pancreas by Studies of Hormone Secretion in Culture and by Xenotransplantation , 1989, Diabetes.

[8]  P. Morris,et al.  Exocrine contamination impairs implantation of pancreatic islets transplanted beneath the kidney capsule. , 1988, The Journal of surgical research.

[9]  D. Eizirik,et al.  Functional restoration of cultured mouse pancreatic islets after in vitro exposure to alloxan. , 1988, Pharmacology & toxicology.

[10]  Y. Ariav,et al.  Monolayer culture of adult rat pancreatic islets on extracellular matrix: long term maintenance of differentiated B-cell function. , 1988, Endocrinology.

[11]  S. Toda,et al.  Unilocular fat cells in three-dimensional collagen gel matrix culture. , 1988, Journal of lipid research.

[12]  P. Lacy,et al.  Low-temperature culture of human islets isolated by the distention method and purified with Ficoll or Percoll gradients. , 1987, Surgery.

[13]  S. Lucke,et al.  The long-term culture of encapsulated pancreatic islets. , 2009, Experimental and clinical endocrinology.

[14]  J. McPherson,et al.  The human immune response to reconstituted bovine collagen. , 1986, Journal of immunology.

[15]  K. Hamaguchi,et al.  Microencapsulation of pancreatic islets. A technique and its application to culture and transplantation. , 1986, Diabetes research and clinical practice.

[16]  A. Monaco,et al.  An improved method for isolation of mouse pancreatic islets. , 1985, Transplantation.

[17]  L. Harrison,et al.  Adult human pancreatic islet cells in tissue culture: function and immunoreactivity. , 1985, The Journal of clinical endocrinology and metabolism.

[18]  M. Jaffrin,et al.  A U-Shaped Bioartificial Pancreas with Rapid Glucose-Insulin Kinetics: In Vitro Evaluation and Kinetic Modelling , 1984, Diabetes.

[19]  W. Silvers,et al.  Influence of culturing on the functioning of major-histocompatibility-complex-compatible and incompatible islet grafts in diabetic mice. , 1983, Transplantation.

[20]  L. Orci,et al.  Collagen matrix promotes reorganization of pancreatic endocrine cell monolayers into islet-like organoids , 1983, The Journal of cell biology.

[21]  D Pipeleers,et al.  Glucose-induced insulin release depends on functional cooperation between islet cells. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[22]  R. Alejandro,et al.  Tissue Culture Reduces la Antigen-bearing Cells in Rat Islets and Prolongs Islet Allograft Survival , 1982, Diabetes.

[23]  P. Lacy,et al.  Prolongation of murine islet allograft survival by pretreatment of islets with antibody directed to Ia determinants. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[24]  K. Lafferty,et al.  Reversal of diabetes by allogenic islet transplantation without immunosuppression. , 1980, The Australian journal of experimental biology and medical science.

[25]  E Bell,et al.  Production of a tissue-like structure by contraction of collagen lattices by human fibroblasts of different proliferative potential in vitro. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[26]  P. Lacy,et al.  PROLONGATION OF ISLET ALLOGRAFT SURVIVAL , 1979, Transplantation.

[27]  M. Kedinger,et al.  In vitro culture reduces immunogenicity of pancreatic endocrine islets , 1977, Nature.