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Diabetologia. 1994 Jan;37(1):15-21.

Effect of nutrients, hormones and serum on survival of rat islet beta cells in culture.

Author information

1
Department of Metabolism and Endocrinology, Vrije Universiteit Brussel, Belgium.

Abstract

This study quantifies the survival of purified single rat beta cells under different culture conditions. Less than 10% of the cells survive 9 days of culture in Ham's F10 medium without supplements. Addition of fetal calf serum (5%) increases cell survival to 54% in the absence and to 78% in the presence of isobutylmethylxanthine (50 mumol/I). The effect of serum is explained, at least partly, by the presence of albumin and of low molecular weight constituents. In serum-free Ham's F10 with 50 mumol/l isobutylmethylxanthine, 75% of cells survive after the addition of bovine serum albumin (1%) and of ultroser (0.2%), a commercial serum substitute. Survival of at least 75% of cells is also maintained in Ham's F10 with isobutylmethylxanthine plus albumin, and supplemented by metabolizable nutrients or by the peptides glucagon (10(-8) mol/l) or growth hormone (1 micrograms/ml) plus insulin like growth factor-I (50 ng/ml). D-Glucose increases beta-cell survival in a dose-dependent manner up to 10 mmol/l; a beneficial effect is also observed with other metabolizable compounds (leucine and glutamine) but not with non-metabolizable monosaccharides. Glucose-induced survival of islet beta cells can be attributed to its dose-dependent recruitment of cells into metabolic activities; however, a 9-day exposure to excessively high nutrient concentrations (> 20 mmol/l glucose) is deleterious to the cells. These results define culture media, with or without serum, wherein at least 75% of single rat islet beta cells can survive for a minimum of 9 days. This will allow for studies on beta-cell toxic conditions and potentially protective agents.(ABSTRACT TRUNCATED AT 250 WORDS).

PMID:
7512059
DOI:
10.1007/bf00428772
[Indexed for MEDLINE]

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