title: Effect of nutrients, hormones and serum on survival of rat islet beta cells in culture.
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)
authors.affiliation: Department of Metabolism and Endocrinology, Vrije Universiteit Brussel, Belgium.
authors.forename: Z
authors.initials: Z
authors.lastname: Ling
authors.forename: J C
authors.initials: JC
authors.lastname: Hannaert
authors.forename: D
authors.initials: D
authors.lastname: Pipeleers
mesh_terms: 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
chemical_list: 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