Culture and Function in Vitro

Abstract

Human fetal pancreas, obtained at prostaglandin-in-duced legal abortions, was maintained in organ culture for 6–14 days in medium TCM 199 (5.5 mM glucose) or RPMI 1640 (11.1 mM glucose) supplemented with 20% calf serum and antibiotics. Light microscopic examination indicated that the cultured explants were composed of fibrous tissue and well-preserved ducts and islets, whereas acinar cells had disappeared. The hormone content (insulin, C-peptide, glucagon, somatostatin) per milligram wet pancreatic weight increased 10–35 times during the culture period. In static incubations of cultured explants there was a poor insulin response to glucose (16.7 mM). By contrast, insulin release was markedly stimulated in the presence of both glucose (16.7 mM) and theophylline (10 mM). The maximal rate of insulin release in response to glucose plus theophylline was observed after 11 days of culture. When pancreatic explants were incubated in a perifusion system after 7 days in culture, 25% of the fetuses showed a low and monophasic insulin response to glucose alone (15 mM), whereas the remaining ones failed to respond. When perifusions were performed with glucose (15 mM) plus theophylline (5 mM), the insulin response was high in about 25% of the fetuses, intermediate in 40%, and low in 35%. Neither in the static incubations nor in the perifusions was there any clear correlation between the rates of insulin release and the tissue culture medium used. It is concluded that human fetal B cells may retain viability in organ culture for more than a week. The fetal pancreatic material prepared and stored by this means may be used in attempts to cure human diabetes by transplantation.