Human Pancreatic Cells in Continuous Culture

DESCRIPTION (provided by applicant): The proposed program as outlined should result in further important characterization and development of immortalized or restored cells of human pancreatic origin. Markers of differentiation and function, mainly insulin production and responsiveness to high glucose loads, will be used to identify populations of higher interest. These populations will be selected for transfection with the hTERT gene, and immortalized. Telomere-enhanced component strains will be isolated, identified, expanded, and further characterized. Telomerase activities and telomere length assays will be performed. To assess the functional consequences of the constitutive or periodic expression of hTERT, a series of structural and phenotypic tests will be performed. Positive controls will consist of related lines normal or immortalized in similar fashion but with the HPV16 E6/E7 genes (on hand). The functional integrity of our lines will be assessed in vivo, and our shielding strategy can be tested in NOD mice with autoimmune prone mechanisms in place. Thus, our cell line generation and engineering strategy should provide sets of potentially therapeutic populations for further developmental and transplantation research.