›› 2011, Vol. 31 ›› Issue (8): 1078-.doi: 10.3969/j.issn.1674-8115.2011.08.006

• Monographic report (Ophthalmology and Visual Sciences) • Previous Articles     Next Articles

Adeno-associated virus mediated transduction of pigment epithelium-derived factor to human iris pigment epithelial cells in vitro

ZHAO Chun1,2, WANG Fang2, JI Jing1, SHI Wo-dong1, FAN Xian-qun1, LUO Min1   

  1. 1.Department of Ophthalmology, the Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, China；2.Department of Ophthalmology, the Tenth People's Hospital, Tongji University, Shanghai 200072, China
  • Online:2011-08-28 Published:2011-08-29
  • Supported by:

    Shanghai Municipal Health Bureau Foundation, 2006028

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Abstract:

Objective To investigate the feasibility of recombinant adeno-associated virus (AAV) mediated transduction of pigment epithelium-derived factor (PEDF) to cultured human iris pigment epithelial (IPE) cells in vitro to obtain the transgenic cells with high expression of PEDF. Methods Recombinant AAV vector carrying PEDF gene (AAV-PEDF) was constructed, and human IPE cells were cultured in vitro and infected with condensed virus. The expression of green fluorescent protein in cells was observed under fluorescence microscopy, the transfection efficiency was detected by flow cytometry, and the expression of PEDF in IPE cells was detected by RT-PCR and Western blotting. Results The proliferation of IPE cells was normal after infection with AAV-PEDF. The expression of PEDF was observed under fluorescence microscopy and detected by RT-PCR and Western blotting. When the multiplicity of infection (MOI) was 20, the transduction efficiency was 64.22%. Conclusion AAV-PEDF can effectively transfect IPE cells cultured in vitro, and the transgenic cells have a high expression of PEDF.

Key words: adeno-associated virus, pigment epithelium-derived factor, iris pigment epithelial cell, transduction