Effect of recombinant adenovirus vector carrying human KGF gene on the proliferation of human epidermal stem cells
LI Xin-Beng, BAI Li-Meng, BAI Wen-Fang, TUN Zhi-Dong, TUN Bo, ZHANG Ming-Sheng
Chinese Journal of Clinical Anatomy ›› 2011, Vol. 29 ›› Issue (5) : 561-564.
Effect of recombinant adenovirus vector carrying human KGF gene on the proliferation of human epidermal stem cells
Objective To construct a recombinant adenovirus vector carrying human keratinocyte growth factor(KGF) gene, and investigate its proliferative effect on the transferred primary human epidermal stem cells. Methods RT-PCR was used to amplify the sepuence of human keratinocyte growth factor. Recombinant adenoviral vector containing the sequence was prepared using pAD-easy-I system. The primary human epidermal stem cells were transfected with the recombinant virus vector. Under the fluorescence microscope, the transfection efficiency was determined. MTT assay was used to analyze the proliferation of hESCs. Cell cycle of hESCs was evaluated by flow cytometry. Results KGF gene was amplified as expected. The recombinant adenoviral plasmid could be digested with Pac I to produce a fragment of 4.5kbp. The transfection efficiency of the recombinant virus at the MOI of 50 was 90%. The proliferation of transgenic cells significantly increased compared to that of controls, evaluated by MTT and the ratio of G2/M cell of the transgenic group. Conclusions The recombinant adenovirus vector containing KGF gene was successfully constructed. The KGF transgenic hESCs showed favourable proliferative potential.
Keratinocyte growth factor / Human epidermal stem cells / Proliferation / Adenovirus
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