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Pancreatic Beta Cell Culture: Preparation of Purified Monolayers

Elliott P. Joslin Research Laboratory, Joslin Diabetes Foundation, Inc., The Harvard Medical School and The Peter Bent Brigham Hospital Boston, Massachusetts 02215

Abstract

Procedures were developed for preparing partially purified beta cell monolayer cultures. Neonatal rat pancreases were dissociated with a trypsin-collagenase solution. Beta cells were separated from denser acinar cells by centrifuging the initial suspension in a two layer discontinuous Ficoll gradient (20, 25%). Resultant cultures, highly enriched in beta cells, were further purified by incubation with cystine-free medium. This caused necrosis of the majority of fibroblastoid cells within two days, while beta cells were considerably less affected. The resultant cultures contained an average of 72% beta cells compared to 10% in untreated control cultures. Insulin release by purified monolayers remained responsive to changes in the glucose concentration of the culture medium. (Endocrinology 96: 637, 1975)

Received September 23, 1974.