Key words Islets, beta cells, culture, glucose, growth hormone.
Springer Online Journal Archives 1860-2000
Summary 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 µmol/l). 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 µmol/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 µg/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. The data also serve as basis for developing media with better survival of beta cells in cultured aggregates. [Diabetologia (1994) 37: 15–21]
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