OBJECTIVE: The aim of this study is to optimize conditions for labeling human periodontal ligament stem cells (PDLSCs) using enhanced green fluorescent protein (eGFP) infected by lentivirus vector and to obtain PDLSCs with high stable expressed eGFP. METHODS: PDLSCs were transfected with eGFP by lentivirus vector for 48 h via different multiplicity of infection (MOI) (25, 50, 100, 200 and 400) and the infection efficiency were analyzed by both fluorescent microscope and flow cytometry. The proliferation rate of infected PDLSCs was evaluated by MTT. The infected PDLSCs were further for detection of pluripotent, differentiation ability and alkaline phosphatase (ALP) expression ability. RESULTS: The infection efficiency for each group were 44.7%, 60.9%, 71.7%, 85.8% and 86.9% respectively. Proliferation of PDLSCs was not affected when MOI was below 200; however, at MOI 400, the proliferation ability was affected compared with control group. The pluripotent and ALP abilities of PDLSCs were not changed by the infection. CONCLUSION: Infection for 48 h at MOI 200 is optimal for labeling PDLSCs with eGFP using lentivirus vector, and the proliferation and differentiation abilities of PDLSCs are not affected obviously.
OBJECTIVE: The aim of this study is to optimize conditions for labeling human periodontal ligament stem cells (PDLSCs) using enhanced green fluorescent protein (eGFP) infected by lentivirus vector and to obtain PDLSCs with high stable expressed eGFP. METHODS: PDLSCs were transfected with eGFP by lentivirus vector for 48 h via different multiplicity of infection (MOI) (25, 50, 100, 200 and 400) and the infection efficiency were analyzed by both fluorescent microscope and flow cytometry. The proliferation rate of infected PDLSCs was evaluated by MTT. The infected PDLSCs were further for detection of pluripotent, differentiation ability and alkaline phosphatase (ALP) expression ability. RESULTS: The infection efficiency for each group were 44.7%, 60.9%, 71.7%, 85.8% and 86.9% respectively. Proliferation of PDLSCs was not affected when MOI was below 200; however, at MOI 400, the proliferation ability was affected compared with control group. The pluripotent and ALP abilities of PDLSCs were not changed by the infection. CONCLUSION:Infection for 48 h at MOI 200 is optimal for labeling PDLSCs with eGFP using lentivirus vector, and the proliferation and differentiation abilities of PDLSCs are not affected obviously.