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Establishment of a long-term stable β-cell line and its application to analyze the effect of Gcg expression on insulin secretion.

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.

journal_name

Sci Rep

journal_title

Scientific reports

authors

Miyazaki S,Tashiro F,Tsuchiya T,Sasaki K,Miyazaki JI

doi

10.1038/s41598-020-79992-7

subject

Has Abstract

pub_date

2021-01-12 00:00:00

pages

477

issue

1

issn

2045-2322

pii

10.1038/s41598-020-79992-7

journal_volume

11

pub_type

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