Electrotransfection by human telomerase reverse transcriptase gene: optimization in precartilagious stem cell culture

doi:10.3969/j.issn.2095-4344.2016.14.020

Abstract

Abstract:

BACKGROUND: The human telomerase reverse transcriptase gene (hTERT) transfected into target cells can play an important role in target cell proliferation and differentiation by increasing telomerase activity and maintaining telomere length.

OBJECTIVE: To explore the effect of hTERT transfection on telomerase activity and biological characteristics of precartilage stem cells culured in vitro.

METHODS: Precartilage stem cells cultured in vitro were subjected to hTERT gene transfection via a retrovirus vector pLXSN. Meanwhile, control and negative control groups were set up. After transfection, TRAP-ELISA assay was used to detect telomerase activity; RT-PCR and western blot employed to detect hTERT mRNA and protein expressions; cell counting kit-8 used to detect cell proliferaiton based on cell growth curve; and flow cytometry adopted to detect cell cycle and distribution.

RESULTS AND CONCLUSION: The telomerase activity was significantly increased at 48 hours after hTERT gene was transfected into the precartilage stem cells. After transfection of hTERT, hTERT mRNA and protein levels were significantly increased, the cell growth rate was significantly increased, the proportion of cells at G₀/G₁ phase was decreased, and the number of S-phased cells increased compared with the control group and negative control group. There were significant differences among the groups (P < 0.05). In conclusion, hTERT transfection via retrovirus vector pLXSN can promote the proliferation of precartilage stem cells in rats by increasing the telomerase activity.
中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Telomerase, Transfection, Chondrocytes, Tissue Engineering

Peng Yi, Qu Jia-fu, Cao Li-hai, Zhao Guo-zhi, Du Xiao-jian. Electrotransfection by human telomerase reverse transcriptase gene: optimization in precartilagious stem cell culture[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(14): 2110-2116.

Figures/Tables 6

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