Morphological and functional characteristics of islets neoformed during tissue culture of fetal rat pancreas

Abstract

Cell suspensions prepared by collagenase digestion of the pancreas of rat fetuses (21.5 days) were cultured for 7-9 days in RPMI medium containing 10 mM glucose. Exocrine cells disappeared rapidly, whereas fibroblasts and endocrine cells proliferated. These latter were first arranged in monolayers but progressively reorganized in neoformed islets essentially composed of B-cells. Total insulin content of the culture dishes increased until day 9, and fractional insulin release was about 20% per day. After 1 week, islets incubated in glucose-free medium released less than 1% of their insulin content over 2 h. Glucose (16.7 mM) caused a slower and weaker (3-fold) stimulation than 10 mM leucine or arginine (3-5-fold). The effects of the three secretagogues were potentiated by theophylline, but only those of glucose and leucine were inhibited by diazoxide. These neoformed islets thus retain a fetal character (relatively low responsiveness to glucose), but the stimulus-specificity of the inhibition by diazoxide is the same as in adult islets. This technique may be useful for studying the mechanisms which govern the organization of pancreatic endocrine cells in islets, and which underlie their functional maturation during the perinatal period.