Establishment of a long-term stable β-cell line and its application to analyze the effect of Gcg expression on insulin secretion.
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| Abstract |
A pancreatic β-cell line MIN6 was previously established in our lab from an insulinoma developed in an IT6 transgenic mouse expressing the SV40 T antigen in β-cells. This cell line has been widely used for in vitro analysis of β-cell function, but tends to lose the mature β-cell features, including glucose-stimulated insulin secretion (GSIS), in long-term culture. The aim of this study was to develop a stable β-cell line that retains the characteristics of mature β-cells. Considering that mice derived from a cross between C3H and C57BL/6 strains are known to exhibit higher insulin secretory capacity than C57BL/6 mice, an IT6 male mouse of this hybrid background was used to isolate insulinomas, which were independently cultured. After 7 months of continuous culturing, we obtained the MIN6-CB4 β-cell line, which stably maintains its GSIS. It has been noted that β-cell lines express the glucagon (Gcg) gene at certain levels. MIN6-CB4 cells were utilized to assess the effects of differential Gcg expression on β-cell function. Our data show the functional importance of Gcg expression and resulting basal activation of the GLP-1 receptor in β-cells. MIN6-CB4 cells can serve as an invaluable tool for studying the regulatory mechanisms of insulin secretion, such as the GLP-1/cAMP signaling, in β-cells.
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| Authors | Satsuki Miyazaki, Fumi Tashiro, Takashi Tsuchiya, Kazuki Sasaki, Jun-Ichi Miyazaki |
| Journal | Scientific reports (Sci Rep) Vol. 11 Issue 1 Pg. 477 (01 12 2021) ISSN: 2045-2322 [Electronic] England |
| PMID | 33436850 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't) |
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