Experimental Study On P63 Gene Transfecting Human Adipose Stem Cells Mediated By Lentivirus

Objective :To make a foundation for further studies on Stem cells differentiating into epidermis cells, the research makes lentivirus as vector, P63 gene was myoblasted into the ADSCs which was cultured in vitro,obtain high expression P63 gene of adipose stem cells.Methods:(1) Get abdominal fat from healthy male under aseptic condition. Separate ADSCs using digestion method, culture in vitro, amplify and store for sparing.(2)Identification of ADSCs: the detection of morphology, multipotent differentiation ability and surface marker.(3) P63 gene myoblasted human adipose derived stem/stromal cells mediated by Lentivirus.(4) The myoblasted cells were detected in PCR and Western Blot.Results:After isolated culture, the obtained cells with spindle-shape were adherent monolayer cells,and were arranged in whirlpool or radiation.The cell growth curve was a typical “S”shape;Oil red O staining was positive, the control group was negative. After ossification induction, alizarin red staining was positive, the control group was negative.Detection of CD14 and CD45 were negative by flow cytometry, CD29、CD44 and CD105 were positive. The myoblasted ADSCs had obviously P63 and keratin expression.Conclusion:Human adipose tissue which acquired through surgical operation contains ADSCs, after culturing in vitro, lentivirus as vector, myoblast P63 gene, cells showed a better ability to epidermal cell differentiation. The resarech has provided reference for the study of ADSCs differentiating into epidermal cell.