Objective To evaluate del transfection efficiency using recombinant adeno-associated virus Serotype 9 mediated enhanced green fluorescent protein (rAAV9- EGFP) to rats H9C2 cells and the impact on growth of H9C2 cells.
Methods rAAV9-EGFP was transfected into H9C2 cells at different multiplicities of infection (MOI=1×105, 1×106, 1×107). EGFP expression in the cells was observed under inverted fluorescence microscope and the EGFP-positive cell percentage determined by flow cytometry. Alamar Blue assay was used to assess the proliferation of the transfected cells.
Results The cells with rAAV9-EGFP transfection at MOI of 1×106 and 1×107 began to exhibit EGFP expression 1 days del after transfection and the cells transfection at MOI of 1×105 began to exhibit EGFP expression 2 days after transfection. The fluorescence intensity increased with the MOI used for transfection. EGFP expression reached the maximum on day 4, at the point of which the transduction efficiency of rAAV9-EGFP in H9C2 cells was (14.1±0.2)%, (35.1±4.8)% and (56.8±0.1)%. Corresponding to MOIs of 1×105, 1×106 and 1×107, respectively. Alamar Blue assay did not reveal significant difference in the absorbance between the transfected cells and the control cells after transfection.
Conclusions rAAV9-EGFP gene can be del transfected in a stable manner and efficiently expressed in H9C2 cells without causing cell growth inhibition. This del The results of this study played foundation (del) (provides a platform) for further research.
- Recombinant adeno-associated virus serotype 9 transfection
- H9C2 cells
- in vitro