Zhai Hui

GW24-e0989 No significant difference of adeno-associated virus serotype 9 transfection efficiency comparison in different cells

Objectives To make a comparison on the efficiency of two kinds of cells for transfecting adeno-associated virus serotype 9 carried enhanced green fluorescence protein, and to demonstrate that adeno-associated virus serotype 9 transfection is feasibility and safety. Methods Human Umbilical Vein Endothelial Cells (HUVECs) and Human Aortic Smooth Muscle Cells (HASMCs) respectively infected with AAV9-eGFP were observed and analysed with fluorescence microscopy and flow cytometry to assess transfection efficiency. Multiplicity of Infection was set to three groups:1*105, 1*106 and 1*107. Non-transfected cells were set as control. Results HUVECs and HASMCs infected with AAV9-eGFP were begin to expressed after transfected 24 hours, and the expression peak appeared in the fourth day in HUVECs, and the fifth day in HASMCs. Cell growth was not affected and cell form was normal in the whole observation period. Multiplicity of infection of 1*105, 1*106 and 1*107 in the fourth day after transfection in HUVECs was 1.4%, 12.3% and 52.8%. And the fifth day in HASMCs was 5.3%, 18.3% and 52.4%. Conclusions There was no obvious differences between HUVECs and HASMCs after transfection in multiplicity of infection of 1*107. This study demonstrates that adeno-associated virus serotype 9 transfection of HUVECs and HASMCs is feasibility and safety.