Maintenance of neonatal rat B cells in glucose-depleted medium: effect of medium with galactose and 2-deoxyglucose.
Monolayer cultures of the pancreas of the neonatal rat were maintained for 7 days in glucose-depleted TCM 199 medium, supplemented with 5.5 mmol galactose/l, with or without 0.1 mmol 2-deoxyglucose/l. Under culture conditions without 2-deoxyglucose, the responsiveness of B cells supported on galactose was totally abolished by day 7 of culture, and islets degenerated. In contrast, the addition of 2-deoxyglucose to the galactose-supplemented medium promoted the survival and the function of B cells even in a glucose-depleted environment and, in addition, yielded the monolayers mostly consisting of endocrine cells by destroying fibroblasts selectively. On day 7 the recovery of insulin in the cells was higher than that of the cells grown in medium with galactose alone (10.7-fold), and than the initial level at day 0 (twofold). Furthermore, the response to an acute challenge with 16.7 mmol glucose/l was 3.3-fold, to 10 mmol leucine/l it was 8.5-fold, and to 10 mmol 2-ketoisocaproate/l it was 10.9-fold above each value observed on day 1. In summary, the above data indicate that morphologically intact, functionally competent endocrine B cells can be grown in medium free from glucose.
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media; D003837:Deoxy Sugars / Q000494:pharmacology*; D003847:Deoxyglucose / Q000494:pharmacology*; D005690:Galactose / Q000494:pharmacology*; D005947:Glucose / Q000494:pharmacology; D007328:Insulin / Q000378:metabolism; D007515:Islets of Langerhans / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D007651:Keto Acids / Q000494:pharmacology; D007930:Leucine / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains
D003470:Culture Media; D003837:Deoxy Sugars; D007328:Insulin; D007651:Keto Acids; C013082:alpha-ketoisocaproic acid; D003847:Deoxyglucose; D007930:Leucine; D005947:Glucose; D005690:Galactose