AAV2-mediated versus AAV2/1-mediated enhanced green fluorescent protein transfection into human skin fibroblast in vitro / 第三军医大学学报

ABSTRACT

Objective To compare the transfection rate of AAV2-EGFP and AAV2/1-EGFP to human skin fibroblast(HFB).Methods Two recombinant adeno-associated viruses(AAV) encoding enhanced green fluorescent protein(EGFP) were constructed and transfected to HFB at multiple of infection MOI ranging from 104 to 106.Twenty-four hours after the infection,the expression rates of EGFP on cultured HFB were assessed by flow cytometry and the transfected cells were observed under fluorescence microscope.The killing effect of the virus on the infected cells was assayed by MTT.Results The transfection efficiency of AAV2-EGFP was increased as MOI increased.When MOI was 104,the transfection efficiency of AAV2-EGFP was 7.68%?1.18%;When MOI was 105,that was 52.12%?1.59%;When MOI was 106,that showed no significant increase.However,the transfection efficiency of AAV2/1-EGFP showed no obvious changes at any MOI.Conclusion AAV2 is more efficient than AAV2/1 in transfecting HFB,but neither AAV has a high transfection efficiency.