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《Journal of Applied Clinical Pediatrics》 2009-09
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Study on Migration of Skin Fibroblasts with Telomerase Reconstitution

TANG Jun,QU Yi,LI Xi-hong,ZHAO Feng-yan,TANG Bin-zhi,MU Dei-zhi(Department of Pediatrics,the Second West China Hospital of Sichuan University,Chengdu 610041,Sichuan Province,China)  
Objective To explore the biocharacteristics of the telomerase-reconstituted fibroblasts,especially the variation of cell migration and the related mechanism behind it.Methods Plasmid containing human telomerase reverse transcriptase(hTERT) gene was transfected through lipofectamine into the human fibroblasts cultured in vitro;positive clones were selected by hygromycin B and resulted in telo-merase reconstituted fibroblasts.Telomerase activity was detected by TRAPEZE Telomerase Detection Kit.The expressions of vimentin and collagen were detected by immonocytochemistry to identify the cell type.Cell migration was evaluated by Boyden Chamber assay.Expression of CXCR4 was detected by RT-PCR and Western blot.Results Introduction of hTERT into fibroblasts resulted in the telomerae reconstitution of the cells.The life-span of telomerase-reconstituted fibroblasts elongated up to 160 passages,and they expressed vimentin and collagen type Ⅰ continuously.When stimulated with a chemokine,CXCL12,the rate of migration was significantly higher in fibroblasts with telomerase-reconstitution than that in fibroblasts without.Furthermore,CXCR4 was upregulated in the telomerase-reconstituted fibroblasts.Conclusions Telomerase reconstitution enhances fibroblast migration through activation of CXCL12/CXCR4 axis.The finding that fibroblasts with telomerase reconstitution has enhanced migration may have broad implications,especially when these fibroblasts are used in cell therapy.
【Fund】: 国家自然科学基金项目资助(30770748);; 四川省科技厅基金项目资助(08ZQ026-069 07JY029-067);; 四川省卫生厅基金项目资助(070164)
【CateGory Index】: R746
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