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《Chinese Journal of Anatomy》 2006-03
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Expression of exogenous insulin-like growth factor 1 gene in bone marrow stromal cells and its effects on cell biological behavior in rat

WU Yan,SONG Jian,WAN Yu,HU Cheng-Jun,LIU Wei-Xin,WANG Qiao(Faculty of Anatomy and Embryology,School of Medicine,Wuhan University,Wuhan 430071,China)  
Objective: To investigate the expression of exogenous hIGF-1 gene in the bone marrow stromal cells(BMSC) and its effects on cell growth and differentiation.Methods: A retrovirous expression vector(pBabe-puro/hIGF-1) was constructed and transfected into cultured rat BMSC.After selection with puromycin,the expression of hIGF-1 in transfected BMSC was examined with RT-PCR assay and ELISA.The growth and natural differentiation of the cells were investigated by MTT assay,and methylene blue,AP and Sudan-Ⅲ staining.A further comparison between the transfected BMSC with or without puromycin selection was conducted to investigate the effects of puromycin on the cell self-renewal.Results: In transfected and puromycin-selected BMSC the expression of hIGF-1 was higher than that in the transfected but not selected cells(10.7-folds: 6.7-folds).The unselected BMSC proliferated more quickly than the control cells,but after selection a weakened ability of growth and colony formation,with enhanced differentiation towards the adipocyte-and osteoblast-like cells were detected.Conclusion: Overexpression of IGF-1 can be induced in BMSC through in vitro gene modification,which promotes the cell proliferation and natural differentiation.However,puromycin selection may have a negative effect on the cell self-renewal.
【Fund】: 湖北省自然科学基金(2002AB00150)
【CateGory Index】: R329
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