Abstract:

Objective

To compare the transfection rate of rAAV2-EGFP and rAAV2/1-EGFP to human pulp cells.

Methods

Two recombinant adeno-associated viruses （AAV）encoding enhanced gene fluorescent protein （EGFP） were constructed and transfected to human dental pulp cells at varying multiple of infection （MOI） of 10⁴，10⁵ and 5×10⁵. Twenty four hours after the infection， the expression rates of EGFP on cultured human dental pulp cells were assessed by flow cytometry and the transfected cells were observed under fluorescence microscope.The proliferation rates of the infected cells were assayed by MTT.

Results

The transfection efficiency of rAAV2-EGFP was increased along with MOI. When MOI was 10⁵， the transfection efficiency of rAAV2-EGFP was 76.9%， while the transfection efficiency of rAAV2/1-EGFP was 35.8%. Furthermore， the transfection efficiency of rAAV2-EGFP and rAAV2/1-EGFP showed no significant influence on the cell activity.

Conclusion

rAAV2-EGFP is more efficient than rAAV2/1-EGFP in transfecting human dental pulp cells.

Key words: Adeno-associated virus, Human dental pulp cell, Fibroblast, Tansfection