Skip to Main content Skip to Navigation
Journal articles

A human beta cell line with drug inducible excision of immortalizing transgenes.

Abstract : Access to immortalized human pancreatic beta cell lines that are phenotypically close to genuine adult beta cells, represent a major tool to better understand human beta cell physiology and develop new therapeutics for Diabetes. Here we derived a new conditionally immortalized human beta cell line, EndoC-βH3 in which immortalizing transgene can be efficiently removed by simple addition of tamoxifen. METHODS: We used lentiviral mediated gene transfer to stably integrate a tamoxifen inducible form of CRE (CRE-ERT2) into the recently developed conditionally immortalized EndoC βH2 line. The resulting EndoC-βH3 line was characterized before and after tamoxifen treatment for cell proliferation, insulin content and insulin secretion. RESULTS: We showed that EndoC-βH3 expressing CRE-ERT2 can be massively amplified in culture. We established an optimized tamoxifen treatment to efficiently excise the immortalizing transgenes resulting in proliferation arrest. In addition, insulin expression raised by 12 fold and insulin content increased by 23 fold reaching 2 μg of insulin per million cells. Such massive increase was accompanied by enhanced insulin secretion upon glucose stimulation. We further observed that tamoxifen treated cells maintained a stable function for 5 weeks in culture. CONCLUSIONS: EndoC βH3 cell line represents a powerful tool that allows, using a simple and efficient procedure, the massive production of functional non-proliferative human beta cells. Such cells are close to genuine human beta cells and maintain a stable phenotype for 5 weeks in culture.
Document type :
Journal articles
Complete list of metadatas

Cited literature [30 references]  Display  Hide  Download

https://www.hal.inserm.fr/inserm-01298792
Contributor : Mireille Bos <>
Submitted on : Wednesday, April 6, 2016 - 4:11:47 PM
Last modification on : Thursday, August 20, 2020 - 3:05:27 AM
Long-term archiving on: : Monday, November 14, 2016 - 5:55:06 PM

File

1-s2.0-S2212877815001787-main....
Publisher files allowed on an open archive

Identifiers

Citation

Marion Benazra, Marie-José Lecomte, Claire Colace, Andreas Müller, Cécile Machado, et al.. A human beta cell line with drug inducible excision of immortalizing transgenes.. Molecular metabolism, Elsevier, 2015, 4 (12), pp.916-925. ⟨10.1016/j.molmet.2015.09.008⟩. ⟨inserm-01298792⟩

Share

Metrics

Record views

647

Files downloads

1450