[Recombinant adeno-associated virus serotype 9 transfection of rats H9C2 cells in vitro].
OBJECTIVE
To evaluate the 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 x 10(5), 1 x 10(6), 1 x 10(7)). 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 x 10(6) and 1 x 10(7) began to exhibit EGFP expression 1 days after transfection and the cells transfection at MOI of 1 x 10(5) 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 x 10(5), 1 x 10(6) and 1 x 10(7), 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 stably and efficiently expressed in H9C2 cells without causing cell growth inhibition.This study played foundation for further research.
Gao
Xia
X
Department of Cardiology, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, China. gaoxia999@gmail.com
Ma
Yi-tong
YT
Yang
Yi-ning
YN
Xiang
Yang
Y
Chen
Bang-dang
BD
Liu
Fen
F
DU
Lei
L
D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation; D000229:Dependovirus / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D051381:Rats; D014162:Transfection
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins