Human Pancreatic Islet Cell Clones Secreting Insulin, Glucagon and Somatostatin: Immunocytochemical and Functional Studies

Human pancreatic A-, B- and D-cell clonal strains, named JHPG-1, JHPI-1 and JHPS-1, were established successfully from adult and fetal pancreata by the single cell plating feeder layer method using a modified Rose's chamber. This is the first time that insulin-, glucagon- and somatostatin-producing clonal strains have been separated into continuous clonal cell lines. The cultured cells are epithelial in nature, free of fibroblast contamination, and can be cloned. Under the phase-contrast microscope, the B-cell clone (JHPI-1) was generally oval or round in shape, the A-cell clone (JHPG-1) was bipolar, and the D-cell clone (JHPS-1) was nerve-like with cytoplasmic processes. By the use of immunocytochemical techniques, insulin-, glucagon- and somatostatin-like immunoreactivities were detected in each clone respectively. By radioimmunoassay it was revealed that each clone produced a single pancreatic hormone. The B-cell clone especially, was found to secrete insulin amply and continuously, for over 150 days. The glucagon release responses by the A-cell clone to insulin and glucose were also studied. The clonal strains obtained in this study provide useful systems for the investigation of the cell-biological aspects of human islet cells in vitro.