Combined introduction of Bmi-1 and hTERT immortalizes human adipose tissue-derived stromal cells with low risk of transformation

Biochemical and Biophysical Research Communications
Péter TátraiKatalin Német

Abstract

Adipose tissue-derived stromal cells (ASCs) are increasingly being studied for their usefulness in regenerative medicine. However, limited life span and donor-dependent variation of primary cells such as ASCs present major hurdles to controlled and reproducible experiments. We therefore aimed to establish immortalized ASC cell lines that provide steady supply of homogeneous cells for in vitro work while retain essential features of primary cells. To this end, combinations of human telomerase reverse transcriptase (hTERT), murine Bmi-1, and SV40 large T antigen (SV40T) were introduced by lentiviral transduction into ASCs. The resulting cell lines ASC(hTERT), ASC(Bmi-1), ASC(Bmi-1+hTERT) and ASC(SV40T+hTERT) were tested for transgene expression, telomerase activity, surface immunomarkers, proliferation, osteogenic and adipogenic differentiation, karyotype, tumorigenicity, and cellular senescence. All cell lines have maintained expression of characteristic surface immunomarkers, and none was tumorigenic. However, ASC(Bmi-1) had limited replicative potential, while the rapidly proliferating ASC(SV40T+hTERT) acquired chromosomal aberrations, departed from MSC phenotype, and lost differentiation capacity. ASC(hTERT) and ASC(hTERT+Bmi...Continue Reading

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Citations

Mar 28, 2013·Evidence-based Complementary and Alternative Medicine : ECAM·Deep KwatraShrikant Anant
Aug 17, 2018·Experimental Biology and Medicine·Dóra MihályZoltán Sápi
Feb 11, 2019·Biotechnology and Bioengineering·Janina BurkCornelia Kasper
Aug 8, 2018·International Journal of Molecular Medicine·Shu-Yu FangYang Bi
Apr 7, 2020·Cell Transplantation·Jia-Jie TanXiang-Ping Li
Dec 19, 2019·International Journal of Molecular Sciences·M Piñeiro-RamilS M Díaz-Prado

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