WANG Ji-dong1, MA Xiao-hui2, LAN Fen3, et al. Lenti-eGFP transfection into rabbit corneal epithelial cells in vitro. [J]. Journal of Chongqing Medical University 36(12):1412-1415(2011)
WANG Ji-dong1, MA Xiao-hui2, LAN Fen3, et al. Lenti-eGFP transfection into rabbit corneal epithelial cells in vitro. [J]. Journal of Chongqing Medical University 36(12):1412-1415(2011)DOI:
Lenti-eGFP transfection into rabbit corneal epithelial cells in vitro
Objective:To observe the influence on cell physiological function after lentivirus vectors mediated enhanced green fluorescence protein(eGFP) gene transfection into rabbit corneal epithelial cells in vitro.Methods:The primary and passage rabbit corneal epithelial cells were cultured and the cells were transfected by lentivirus vectors mediated eGFP gene(lenti-eGFP) with the optimum multiplicity of Infection(MOI)=100.After successful transfection
the expression of eGFP was examined through the inverted fluorescence microscope.The cellular morphology and ultra microstructures of the epithelial cells were observed by HE stain and transmission electron microscope.Furthermore
we calculated the proliferation rate of the normal(control) and transfected cells on 2
3 d and 4 d since the culture.The apoptosis rate of both transfection cells and control cells was counted by Flow cytometer(FCM).Results:The eGFP began to express in the cells and was observed by the inverted fluorescence microscope 48 h after transfection and was still seen after the cells were subcultured.At MOI=100
the cellular morphology and ultra microstructure of infected cells were not changed significantly compared with the normal epithelial cells.The cell proliferation rate for the two groups on 2
3 d and 4 d since the culture was 1.07±0.14 vs 1.04±0.19
0.52±0.06 vs 0.53±0.05 and 0.23±0.03 vs 0.21±0.04 respectively.There was no significant difference in the cell proliferation rate between infected cells and control cells(P>0.05).The outcome of apoptosis rate(0.645±0.040 vs 0.650±0.030) suggested no significant difference in both normal and infected cells(P>0.05).Conclusion:Lenti-eGFP could transfect the rabbit epithelial cells ex vivo efficiently and the influence on the cells needs further discussion.
关键词
慢病毒载体基因转染增强型绿色荧光蛋白角膜上皮细胞实验
Keywords
lentivirus vectorsgene transfectionenhanced green fluorescence proteincorneal epithelial cellcell experiment