Experimental research of enhanced green fluorescent protein gene transfection into mesenchymal stem cells in a rabbit

Objective The establishment of methods of Enhanced Green fluorescent Protein(EGFP) labled mesenchymal stem cells(MSCs),and the study of differentiation of EGFP labled MSCs.Methods MSCs were Isolated and cultured by density gradient centrifugation and adhesive culture methods,and transfected with AdV vector with EGFP.The transient expression of EGFP was observed by fluorescence microscope and transfection efficiency was calculated.Also,the differentiation of labled MSCs into endothelial cell was detected.Results①About 80%～90%MSCs were fused on day 16～20, which had diverse configurations of spindle,polygon and fusiform under phase-contrast microscope.Subcultured MSCs gained uniform spindle- shaped in appearance and showed actively proliferative capacity;②The expression of EGFP began in 24 hours after transferring,reached maximum in 4～5 days,then decreased,no EGFP was found in 4 weeks.③Transfer efficiency was about 60%;④Endothelial cells differentiated from induced MSCs show slabstone-like line and express CD31 antigen.Conclusions MSCs may be effectively transfected by Ad-CMV-EGFP and express EGFP,and in which the transfection efficiency is relatively high and not affect the differentiation of MSCs,so MSCs labled with Ad-CMV-EGFP is a good method.