Development of a conditionally immortalized human pancreatic beta cell line

作者：Scharfmann Raphael^*; Pechberty Severine; Hazhouz Asmine; von Buelow Manon; Bricout Neveu Emilie; Grenier Godard Maud; Guez Fanny; Rachdi Latif; Lohmann Matthias; Czernichow Paul; Ravassard Philippe

来源：Journal of Clinical Investigation, 2014, 124(5): 2087-2098.

DOI：10.1172/JCI72674

Diabetic patients exhibit a reduction in beta cells, which secrete insulin to help regulate glucose homeostasis; however, little is known about the factors that regulate proliferation of these cells in human pancreas. Access to primary human beta cells is limited and a challenge for both functional studies and drug discovery progress. We previously reported the generation of a human beta cell line (EndoC-beta H1) that was generated from human fetal pancreas by targeted oncogenesis followed by in vivo cell differentiation in mice. EncloC-beta H1 cells display many functional properties of adult beta cells, including expression of beta cell markers and insulin secretion following glucose stimulation; however, unlike primary beta cells, EndoC-(beta H1 cells continuously proliferate. Here, we devised a strategy to generate conditionally immortalized human beta cell lines based on Cre-mediated excision of the immortalizing transgenes. The resulting cell line (EndoC-beta H2) could be massively amplified in vitro. After expansion, transgenes were efficiently excised upon Cre expression, leading to an arrest of cell proliferation and pronounced enhancement of beta cell-specific features such as insulin expression, content, and secretion. Our data indicate that excised EndoC-beta H2 cells are highly representative of human beta cells and should be a valuable tool for further analysis of human beta cells.

• 出版日期2014-5