Culture of endocrine pancreatic cells in protein-free, chemically defined media.

Cell suspensions prepared by collagenase digestion of pancreata obtained from 21.5-d-old rat fetuses were preincubated in RPMI medium containing 10% fetal bovine serum (FBS), to ensure cell adhesion. Twenty hours later, this medium was replaced by a chemically defined medium.

In vitro cytodifferentiation of perinatal rat islet cells within a tridimensional matrix of collagen.

Cell suspensions prepared by collagenase digestion of pancreases obtained from rat fetuses (21.5 d old) and newborns (2.5 d old) were mixed with a collagen solution and inoculated on a collagen base layer.

Cell interactions during the in vitro neoformation of fetal rat pancreatic islets.

As shown by scanning electron microscopy, transmission electron microscopy and membrane labeling analysis, the in vitro neoformation of rat pancreatic islets arose from two main processes: a budding from explants containing duct cells, and a competition between endocrine monolayers and fibroblasts on the culture substratum. The stronger cytoskeleton of fibroblasts and their higher adhesive properties, probably related to their more homogeneous distribution of membrane charges, may explain the spherization of the islets. The pure endocrine cell population of neoformed islets was composed mainly of insulin-secreting cells, and the other types of endocrine cells were distributed in the periphery.