关键词:  , KGF, 表皮干细胞, 增殖, 腺病毒

Abstract:

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.

Key words: Keratinocyte growth factor, Human epidermal stem cells, Proliferation, Adenovirus