Comparative functional study of clonal insulin-secreting cells cultured in five commercially available tissue culture media

M Hamid, Janie McCluskey, Neville McClenaghan, Peter Flatt

Research output: Contribution to journalArticle

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Abstract

The electrofusion-derived rat insulin-secreting cell line BRIN-BD11 was cultured in five different commercially available media to determine the optimum medium for the in vitro maintenance of such clonal cell lines. Cells were cultured in RPMI-1640. DMEM, McCOY'S, F-12K, or MEM culture medium supplemented with 10% (v/v) fetal bovine serum and antibiotics (100 U/ml penicillin and 0.1 g/L streptomycin). Insulin secretion studies performed after 10 days revealed RPMI-1640 to be the best performing medium in terms of insulin secretory responsiveness to a range of stimuli including glucose, L-alanine, L-arginine, carbachol, and glibenclamide. Insulin release was significantly decreased (p <0.01 to p <0.05) in all other media compared to RPMI-16-10. Only the cells cultured in RPMI-1640 and DMEM showed a significant glucose-induced insulin secretory response (p <0.01 and p <0.05). McCOY'S gave the next best result followed by F-12K and MEM. After the 10-day culture period, the highest insulin content was found in cells cultured in RPMI-1640 and DMEM with significantly lower levels of insulin in cells cultured in McCOY'S, F-12K, and MEM (p <0.01 to p <0.001). RPMI-1640 was used for further studies to investigate the effects of 5.6-16.7 mmol/L glucose in culture on the secretory responsiveness of BRIN-BD11 cells. Significant responses to a number of nonglucidic secretagogues were seen following culture at 5.6 and 16.7 mmol/L glucose, although responsiveness was less than after culture with 11.1 mmol/L glucose. At 16.7 mmol/L glucose culture, glucose-stimulated insulin release was abolished.
LanguageEnglish
Pages153-159
JournalCell Transplantation
Volume10
Issue number2
Publication statusPublished - Mar 2001

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Insulin-Secreting Cells
Culture Media
Insulin
Glucose
Cultured Cells
Cell Line
Glyburide
Carbachol
Streptomycin
Penicillins
Alanine
Arginine
Maintenance
Anti-Bacterial Agents
Serum

Cite this

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title = "Comparative functional study of clonal insulin-secreting cells cultured in five commercially available tissue culture media",
abstract = "The electrofusion-derived rat insulin-secreting cell line BRIN-BD11 was cultured in five different commercially available media to determine the optimum medium for the in vitro maintenance of such clonal cell lines. Cells were cultured in RPMI-1640. DMEM, McCOY'S, F-12K, or MEM culture medium supplemented with 10{\%} (v/v) fetal bovine serum and antibiotics (100 U/ml penicillin and 0.1 g/L streptomycin). Insulin secretion studies performed after 10 days revealed RPMI-1640 to be the best performing medium in terms of insulin secretory responsiveness to a range of stimuli including glucose, L-alanine, L-arginine, carbachol, and glibenclamide. Insulin release was significantly decreased (p <0.01 to p <0.05) in all other media compared to RPMI-16-10. Only the cells cultured in RPMI-1640 and DMEM showed a significant glucose-induced insulin secretory response (p <0.01 and p <0.05). McCOY'S gave the next best result followed by F-12K and MEM. After the 10-day culture period, the highest insulin content was found in cells cultured in RPMI-1640 and DMEM with significantly lower levels of insulin in cells cultured in McCOY'S, F-12K, and MEM (p <0.01 to p <0.001). RPMI-1640 was used for further studies to investigate the effects of 5.6-16.7 mmol/L glucose in culture on the secretory responsiveness of BRIN-BD11 cells. Significant responses to a number of nonglucidic secretagogues were seen following culture at 5.6 and 16.7 mmol/L glucose, although responsiveness was less than after culture with 11.1 mmol/L glucose. At 16.7 mmol/L glucose culture, glucose-stimulated insulin release was abolished.",
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Comparative functional study of clonal insulin-secreting cells cultured in five commercially available tissue culture media. / Hamid, M; McCluskey, Janie; McClenaghan, Neville; Flatt, Peter.

In: Cell Transplantation, Vol. 10, No. 2, 03.2001, p. 153-159.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Comparative functional study of clonal insulin-secreting cells cultured in five commercially available tissue culture media

AU - Hamid, M

AU - McCluskey, Janie

AU - McClenaghan, Neville

AU - Flatt, Peter

PY - 2001/3

Y1 - 2001/3

N2 - The electrofusion-derived rat insulin-secreting cell line BRIN-BD11 was cultured in five different commercially available media to determine the optimum medium for the in vitro maintenance of such clonal cell lines. Cells were cultured in RPMI-1640. DMEM, McCOY'S, F-12K, or MEM culture medium supplemented with 10% (v/v) fetal bovine serum and antibiotics (100 U/ml penicillin and 0.1 g/L streptomycin). Insulin secretion studies performed after 10 days revealed RPMI-1640 to be the best performing medium in terms of insulin secretory responsiveness to a range of stimuli including glucose, L-alanine, L-arginine, carbachol, and glibenclamide. Insulin release was significantly decreased (p <0.01 to p <0.05) in all other media compared to RPMI-16-10. Only the cells cultured in RPMI-1640 and DMEM showed a significant glucose-induced insulin secretory response (p <0.01 and p <0.05). McCOY'S gave the next best result followed by F-12K and MEM. After the 10-day culture period, the highest insulin content was found in cells cultured in RPMI-1640 and DMEM with significantly lower levels of insulin in cells cultured in McCOY'S, F-12K, and MEM (p <0.01 to p <0.001). RPMI-1640 was used for further studies to investigate the effects of 5.6-16.7 mmol/L glucose in culture on the secretory responsiveness of BRIN-BD11 cells. Significant responses to a number of nonglucidic secretagogues were seen following culture at 5.6 and 16.7 mmol/L glucose, although responsiveness was less than after culture with 11.1 mmol/L glucose. At 16.7 mmol/L glucose culture, glucose-stimulated insulin release was abolished.

AB - The electrofusion-derived rat insulin-secreting cell line BRIN-BD11 was cultured in five different commercially available media to determine the optimum medium for the in vitro maintenance of such clonal cell lines. Cells were cultured in RPMI-1640. DMEM, McCOY'S, F-12K, or MEM culture medium supplemented with 10% (v/v) fetal bovine serum and antibiotics (100 U/ml penicillin and 0.1 g/L streptomycin). Insulin secretion studies performed after 10 days revealed RPMI-1640 to be the best performing medium in terms of insulin secretory responsiveness to a range of stimuli including glucose, L-alanine, L-arginine, carbachol, and glibenclamide. Insulin release was significantly decreased (p <0.01 to p <0.05) in all other media compared to RPMI-16-10. Only the cells cultured in RPMI-1640 and DMEM showed a significant glucose-induced insulin secretory response (p <0.01 and p <0.05). McCOY'S gave the next best result followed by F-12K and MEM. After the 10-day culture period, the highest insulin content was found in cells cultured in RPMI-1640 and DMEM with significantly lower levels of insulin in cells cultured in McCOY'S, F-12K, and MEM (p <0.01 to p <0.001). RPMI-1640 was used for further studies to investigate the effects of 5.6-16.7 mmol/L glucose in culture on the secretory responsiveness of BRIN-BD11 cells. Significant responses to a number of nonglucidic secretagogues were seen following culture at 5.6 and 16.7 mmol/L glucose, although responsiveness was less than after culture with 11.1 mmol/L glucose. At 16.7 mmol/L glucose culture, glucose-stimulated insulin release was abolished.

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JO - Cell Transplantation

T2 - Cell Transplantation

JF - Cell Transplantation

SN - 0963-6897

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