Effect of nutrients, hormones and serum on survival of rat islet beta cells in culture.
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)
Ling
Z
Z
Department of Metabolism and Endocrinology, Vrije Universiteit Brussel, Belgium.
Hannaert
J C
JC
Pipeleers
D
D
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000818:Animals; D001769:Blood; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D003470:Culture Media; D016895:Culture Media, Serum-Free; D005934:Glucagon / Q000494:pharmacology; D005947:Glucose / Q000494:pharmacology; D005973:Glutamine / Q000494:pharmacology; D013006:Growth Hormone / Q000494:pharmacology; D006728:Hormones / Q000494:pharmacology*; D007328:Insulin / Q000378:metabolism; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology; D007515:Islets of Langerhans / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D007930:Leucine / Q000494:pharmacology; D008297:Male; D009005:Monosaccharides / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D012710:Serum Albumin, Bovine / Q000494:pharmacology
D003470:Culture Media; D016895:Culture Media, Serum-Free; D006728:Hormones; D007328:Insulin; D009005:Monosaccharides; D005973:Glutamine; D012710:Serum Albumin, Bovine; D007334:Insulin-Like Growth Factor I; D013006:Growth Hormone; D005934:Glucagon; D007930:Leucine; D005947:Glucose; D015056:1-Methyl-3-isobutylxanthine