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Characteristics of BRIN-BG5 and BRIN-BG7, two novel glucose-responsive insulin-secreting cell lines produced by electrofusion

Characteristics of BRIN-BG5 and BRIN-BG7, two novel glucose-responsive insulin-secreting cell... AbstractTwo hybrid insulin-secreting cell lines (BRIN-BG5 and BRIN-BG7) were established by the novel approach of electrofusing RINm5F cells with New England Deaconess Hospital rat pancreatic islet cells. Cells were selected from the fusion mixture on the basis of insulin output. Wells showing five to ten times greater insulin output than parental RINm5F cells were selected, subcultured and cloned. Clonal BRIN-BG5 and BRIN-G7 cells grow as monolayers with epithelial morphology. The differences in doubling time of 28 and 20 h respectively were associated with morphological differences; the growth pattern and insulin content of each cell line remaining stable for over 50 passages. In acute 20-min tests, both cell lines showed peak secretory responses (1·9- and 1·8-fold respectively) to 8·4 mmol/l glucose. Membrane depolarization with 25 mmol/l K+ evoked 3·7- and 3·9-fold increases in insulin output. l-Alanine (10 mmol/l) also served to promote 2·4- and 1·6-fold increases in insulin release respectively. Increasing the Ca2+ concentration from 1·28 to 7·68 mmol/l potentiated this effect by 1·8- and 1·5-fold. Incubation with forskolin (25 μmol/l) or phorbol-12-myristate 13-acetate (10 nmol/l), in the presence of l-alanine, similarly enhanced the secretory effect on BRIN-BG5 and BRIN-BG7 cells by 1·3- to 2·1-fold and 1·2- to 1·5-fold respectively. The presence of a functional glucose-sensing mechanism in both cell lines was confirmed by the demonstration of the glucose transporter GLUT-2 and measurement of glucokinase activity. These functional properties suggest that insulin-secreting BRIN-BG5 and BRIN-BG7 cells represent two useful glucoseresponsive cell lines for future studies of the function of the pancreatic B-cell.Journal of Endocrinology (1996) 148, 409–417 http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Endocrinology Bioscientifica

Characteristics of BRIN-BG5 and BRIN-BG7, two novel glucose-responsive insulin-secreting cell lines produced by electrofusion

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Publisher
Bioscientifica
Copyright
Copyright © 1996 The Authors. All Rights Reserved.
ISSN
0022-0795
eISSN
1479-6805
DOI
10.1677/joe.0.1480409
Publisher site
See Article on Publisher Site

Abstract

AbstractTwo hybrid insulin-secreting cell lines (BRIN-BG5 and BRIN-BG7) were established by the novel approach of electrofusing RINm5F cells with New England Deaconess Hospital rat pancreatic islet cells. Cells were selected from the fusion mixture on the basis of insulin output. Wells showing five to ten times greater insulin output than parental RINm5F cells were selected, subcultured and cloned. Clonal BRIN-BG5 and BRIN-G7 cells grow as monolayers with epithelial morphology. The differences in doubling time of 28 and 20 h respectively were associated with morphological differences; the growth pattern and insulin content of each cell line remaining stable for over 50 passages. In acute 20-min tests, both cell lines showed peak secretory responses (1·9- and 1·8-fold respectively) to 8·4 mmol/l glucose. Membrane depolarization with 25 mmol/l K+ evoked 3·7- and 3·9-fold increases in insulin output. l-Alanine (10 mmol/l) also served to promote 2·4- and 1·6-fold increases in insulin release respectively. Increasing the Ca2+ concentration from 1·28 to 7·68 mmol/l potentiated this effect by 1·8- and 1·5-fold. Incubation with forskolin (25 μmol/l) or phorbol-12-myristate 13-acetate (10 nmol/l), in the presence of l-alanine, similarly enhanced the secretory effect on BRIN-BG5 and BRIN-BG7 cells by 1·3- to 2·1-fold and 1·2- to 1·5-fold respectively. The presence of a functional glucose-sensing mechanism in both cell lines was confirmed by the demonstration of the glucose transporter GLUT-2 and measurement of glucokinase activity. These functional properties suggest that insulin-secreting BRIN-BG5 and BRIN-BG7 cells represent two useful glucoseresponsive cell lines for future studies of the function of the pancreatic B-cell.Journal of Endocrinology (1996) 148, 409–417

Journal

Journal of Endocrinology – Bioscientifica

Published: Mar 1, 1996

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