Abstract: The present invention provides a method of culturing cells on a matrix. The present invention includes islet cells cultured in a 3-D configuration in the presence of a fibrin matrix support. Induction of &bgr;-cell proliferation with HGF/SF is also provided.

Abstract: This invention relates to cell lines, particularly mammalian cell lines, established by transforming the cells with vectors, preferably retroviral vectors, containing two or more oncogenes under the control of one or more inducible promoters and/or genetic elements. Also within the scope of the invention are human cell lines with extended in vitro lifespan, transformed by vectors containing one or more oncogenes under the control of one or more, preferably exogenous, inducible promoters and/or genetic elements. The vectors may additionally contain gene(s) encoding for desired gene product(s). Also disclosed are insulin producing human pancreatic cell lines useful for transplantation into human diabetic patients.

Abstract: This invention pertains to the discovery that inhibition of phosphatidylinositol 3-kinase (PI3K) in human fetal undifferentiated cells induces morphological and functional endocrine differentiation. This is associated with an increase in mRNA levels of insulin, glucagon, and somatostatin, as well as an increase in the insulin protein content and secretion response to secretagogues. Blockade of PI3K also increases the proportion of pluripotent precursor cells coexpressing multiple hormones and the total number of terminally differentiated cells originating from these precursor cells.

Abstract: This invention pertains to the discovery that inhibition of phosphatidylinositol 3-kinase (PI3K) in human fetal undifferentiated cells induces morphological and functional endocrine differentiation. This is associated with an increase in MRNA levels of insulin, glucagon, and somatostatin, as well as an increase in the insulin protein content and secretion response to secretagogues. Blockade of PI3K also increases the proportion of pluripotent precursor cells coexpressing multiple hormones and the total number of terminally differentiated cells originating from these precursor cells. In one embodiment, this invention thus rpov9des methods for endocrine differentiation in cultured cells.

Abstract: This invention relates to cell lines, particularly mammalian cell lines, established by transforming the cells with vectors, preferably retroviral vectors, containing two or more oncogenes under the control of one or more inducible promoters and/or genetic elements. Also within the scope of the invention are human cell lines with extended in vitro lifespan, transformed by vectors containing one or more oncogenes under the control of one or more, preferably exogenous, inducible promoters and/or genetic elements. The vectors may additionally contain gene(s) encoding for desired gene product(s). Also disclosed are insulin producing human pancreatic cell lines useful for transplantation into human diabetic patients.

Abstract: Human pancreatic endocrine cells are proliferated without loss of hormone function in a culture medium containing extracellular matrix from bladder carcinoma cell lines in the substantial absence of hepatocyte growth factor/scatter factor. Proliferation is preferably carried out in the substantial absence of any peptide growth factors and nicotinamide. The cells may be proliferated in a monolayer on a solid substrate. Islets and islet-like cell clusters are proliferated without loss of insulin-secreting function by incubation in a medium containing extracellular matrix from a human bladder carcinoma cell line, preferably cell line ATCC HTB-9.

Abstract: The combination of trehalose and dimethyl sulfoxide is an unusually effective cryoprotectant for islets and islet-like cell clusters, as well as platelets. Islets and ICCs, when cooled through the thermotropic phase transition in the presence of this combination of treatment agents and then returned to physiologic temperature, retain their functionality. Likewise, platelets can be similarly cooled and warmed without undergoing premature activation. In general, trehalose can be incorporated into eukaryotic cells in general by suspending the cells in a trehalose solution and either cooling or warming the solution through the thermotropic transition of the cells.

Abstract: This invention relates to cell lines, particularly mammalian cell lines, established by transforming the cells with vectors, preferably retroviral vectors, containing two or more oncogenes under the control of one or more inducible promoters and/or genetic elements. Also within the scope of the invention are human cell lines with extended in vitro lifespan, transformed by vectors containing one or more oncogenes under the control of one or more, preferably exogenous, inducible promoters and/or genetic elements. The vectors may additionally contain gene(s) encoding for desired gene product(s). Also disclosed are insulin producing human pancreatic cell lines useful for transplantation into human diabetic patients.

Abstract: A method of inducing the proliferation and/or differentiation of human fetal pancreatic cells entails contacting such cells in primary culture with Hepatocyte Growth Factor/Scatter Factor, thereby inducing a proliferation of .beta.-epithelial cells, an increase in the number of .beta.-epithelial cells which form islet-like cell clusters, and an increase in insulin production per cell. The method provides increased numbers of functional islet-like cell clusters for transplantation, for example, into Type 1 diabetic patients. The method can be scaled up so as to provide clinically useful numbers of cells for transplantation.

Abstract: The addition of basic FGF-saporin mitotoxins selectively elminates fibroblastoids from human islets in culture and increases the ability of such islets to release insulin under basal and stimulated conditions. When such islets are attached to an extracellular matrix, in particular BCEM, the proliferation of islet cells is favored, and the ability of islets cultured in such manner to release insulin is further increased. Moreover, supplementation of the culture media with high glucose or insulin further improves the functioning of the human islets, resulting in augmented insulin release. Combinations of such procedures offer a novel approach towards the establishment of viable human islet cell monolayers for clinical and laboratory research.