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Cre-mediated reversible immortalization of human renal proximal tubular epithelial cells

Oncogene volume 23, pages 5950–5957 (2004)Cite this article

Abstract

Primary human renal proximal tubule epithelial cells (RPTECs) are of limited use for basic research and for clinical applications due to their limited lifespan in culture. Here we used two lentivirus vectors carrying the human telomerase (hTERT) and the SV40T antigen (Tag) flanked by loxP sites to reversibly immortalize RPTECs. Transduced RPTEC clones continued to proliferate while retaining biochemical and functional characteristics of primary cells. The clones exhibited contact-inhibited, anchorage- and growth factor-dependent growth and did not form tumors in nude mice, suggesting that the cells were not transformed. Transient Cre expression in these cells led to efficient proviral deletion, upregulation of some renal specific activities, and decreased growth rates. Ultimately, the cells underwent replicative senescence, indicating intact cell cycle control. Thus, reversible immortalization allows the expansion of human RPTECs, leading to large production of RPTECs that retain most tissue-specific properties.

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Acknowledgements

We thank Dr RA Weinberg (MIT) for the hTERT cDNA and Dr B Sauer for the cre gene. This work was supported in part by National Institutes of Health Grant (R21 CA101629).

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  1. Department of Virology, Beckman Research Institute, City of Hope, 1500 E. Duarte Road, Duarte, 91010, CA, USA

    Claudia M Kowolik, Shujian Liang, Ying Yu & Jiing-Kuan Yee

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  1. Claudia M Kowolik
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Correspondence to Jiing-Kuan Yee.

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Kowolik, C., Liang, S., Yu, Y. et al. Cre-mediated reversible immortalization of human renal proximal tubular epithelial cells. Oncogene 23, 5950–5957 (2004). https://doi.org/10.1038/sj.onc.1207801

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