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.