The effect  of retrovirus-mediated hTRT transfection into cultured oral keratinocytes

Shanghai Journal of Stomatology ›› 2014, Vol. 23 ›› Issue (3): 304-307.

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The effect of retrovirus-mediated hTRT transfection into cultured oral keratinocytes

HUANG Ji-yan^1,2, LIU Wei³, ZHOU Zeng-tong¹, ZHOU Hai-wen¹

1.Department of Oral Medicine, Ninth People’s Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Stomatology. Shanghai 200011;
2. Department of Stomatology, Shuguang Hospital of Shanghai Chinese Medicine University. Shanghai 201203;
3.Tissue Engineering Research Center of Shanghai Jiao Tong University, Shanghai Key Laboratory of Tissue Engineering. Shanghai 200011, China

Received:2013-12-09 Online:2014-06-20 Published:2014-09-09

Abstract

Abstract: PURPOSE: Human telomerase reverse transcriptase (hTRT) was transfected into cultured oral keratinocytes (OKC) mediated by pBABE-tert recombined retrovirus to investigate the effect on OKC lifespan. METHODS: pBABE-tert recombined retrovirus loaded with hTRT gene was amplified by transfected PT67 cells, and then transfected into cultured OKC in vitro. The positive clones of OKC were separated by puromycin and subcultured. Telomerase activity was analyzed by telomerase PCR-ELISA and PCR-PAGE. RESULTS: The hTRT positive clones of OKC showed telomerase expression, with extending lifespan to 8-9 passages. CONCLUSIONS: The hTRT transfected OKC can prolong doubly lifespan but not be immortalized, which indicates that cellular immortality mechanism is complicated and multi-controled. Telomerase activity is the key for cell immortalization but not the only impact factor.

Key words: Cell culture, Oral Keratinocytes, Recombined retrovirus, hTRT/hTERT, Gene transfection

CLC Number:

R781.5

HUANG Ji-yan, LIU Wei, ZHOU Zeng-tong, ZHOU Hai-wen. The effect of retrovirus-mediated hTRT transfection into cultured oral keratinocytes[J]. Shanghai Journal of Stomatology, 2014, 23(3): 304-307.

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The effect of retrovirus-mediated hTRT transfection into cultured oral keratinocytes

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