Objectives To approach effect of efficiency and growth with Lentiviral transfection Umbilical cord wharton's jelly-derived mesenchymal stem cells (UW-MSCs) in different multiplicity of infection and time.
Methods Lentiviral with enhanced green fluorescent protein (Lenti-EGFP) infection rat UW-MSCs of culture in vitro, multiplicity of infection (MOI) 0, 5, 10, 50 difference, to observe EGFP disclose 2 days, 7 days, 10 days difference, to detect vis vitaev, generation multiple, cell differentiation information of daughter cell, and appraisal effect of UW-MSCs transfection UW-MSCs. To detect transfection efficiency and fluorescence index number (FI) of Lentiviral transfection UW-MSCs used flow cytometer (FCM).
Results 36 h after transfection Lenti-EGFP to discover green fluorescence, brightness different, green fluorescence brightness strengthen gradually with cultivate cell time to extend, chalk steady state afrer 7 days, vis vitaev, generation multiple, cell differentiation information of MOI difference Lenti-EGFP transfection UW-MSCs do not marked change (p>0.05); at same MOI generation multiple to rise striking of 7 days to compare with 2 days and 10 days to compare with 7 days (p<0.01). Transfection efficiency and fluorescence index number increase significant deviation with increase of multiplicity of infection (0, 5, 10, 50) and cultivate cell times (2 days, 7 days, 10 days) (p<0.05).
Conclusions Lenti-EGFP be able to transfection UW-MSCs valid. But transfection efficiency and fluorescence index number increase significant deviation with increase of multiplicity of infection and cultivate cell time. Lenti-EGFP did not effect vis vitaev, generation multiple and cell differentiation information of UW-MSCs. Lentiviral is a kind of active gene transfer vector to reform UW-MSCs.