Pancreatic beta cell culture: preparation of purified monolayers.
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 discontinous 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.
D000818:Animals; D002469:Cell Separation; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D003470:Culture Media; D003553:Cystine; D005069:Evaluation Studies as Topic; D005947:Glucose / Q000494:pharmacology; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D010179:Pancreas / Q000166:cytology* / Q000378:metabolism; D051381:Rats
D003470:Culture Media; D007328:Insulin; D003553:Cystine; D005947:Glucose