1. Development of a conditionally immortalized human pancreatic β cell line
- Author
-
Scharfmann, Raphael, Pechberty, Severine, Hazhouz, Yasmine, von Bulow, Manon, Bricout-Neveu, Emilie, Grenier-Godard, Maud, Guez, Fanny, Rachdi, Latif, Lohmann, Matthias, Czernichow, Paul, and Ravassard, Philippe
- Subjects
Cells -- Aging ,Cell research ,Pancreatic beta cells -- Physiological aspects ,Health care industry - Abstract
Diabetic patients exhibit a reduction in β 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 β cells is limited and a challenge for both functional studies and drug discovery progress. We previously reported the generation of a human β cell line (EndoC-βH1) that was generated from human fetal pancreas by targeted oncogenesis followed by in vivo cell differentiation in mice. EndoC-βH1 cells display many functional properties of adult β cells, including expression of β cell markers and insulin secretion following glucose stimulation; however, unlike primary β cells, EndoC-βH1 cells continuously proliferate. Here, we devised a strategy to generate conditionally immortalized human β cell lines based on Cre-mediated excision of the immortalizing transgenes. The resulting cell line (EndoC-βH2) 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 β cell-specific features such as insulin expression, content, and secretion. Our data indicate that excised EndoC-βH2 cells are highly representative of human β cells and should be a valuable tool for further analysis of human β cells., Introduction Insulin-producing pancreatic β cells play a central role in glycemic regulation. Such β cells are destroyed in patients with type 1 diabetes, while in type 2 diabetes patients, functional [...]
- Published
- 2014
- Full Text
- View/download PDF