›› 2013, Vol. 33 ›› Issue (4): 439-.doi: 10.3969/j.issn.1674-8115.2013.04.012

• Original article (Basic research) • Previous Articles     Next Articles

Transfection efficiency and cytotoxicity of lentivirus-GFP on rabbit adipose-derived stem cells

LI Hong-song1, XU Chen2,3, ZOU Jun1   

  1. 1.Department of Ophthalmology, the Sixth People´s Hospital, Shanghai Jiaotong University, Shanghai 200233, China； 2.Department of Histology and Embryology, Basic Medical College, Shanghai Jiaotong University, Shanghai 200025, China； 3.Shanghai Key Laboratory of Reproductive Medicine, Shanghai 200025, China
  • Online:2013-04-28 Published:2013-05-03

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Abstract:

Objective To evaluate the transfection efficiency and cytotoxicity of lentivirus containing green fluorescence protein (GFP)(lentivirus-GFP) on rabbit adipose-derived stem cells (rADSCs), and establish a method of cell tracer marker in the study of reconstruction of tissue engineered corneal epithelium. Methods rADSCs were transfected with lentivirus-GFP at different multiplicity of infection (MOI). Cell transfection was observed under fluorescent microscope, and influence of lentivirus transfection on proliferation of rADSCs was detected by MTT. Results rADSCs were successfully transfected at MOI of 50, 100 and 500, and the transfection efficiency could reach more than 50% 120 h later. Lentivirus-GFP had no significant effect on proliferation of rADSCs at MOI of 50 and 100, while the proliferation of rADSCs was significantly inhibited at MOI of 500. Conclusion rADSCs are successfully transfected with lentivirus-GFP, which can be used in the study of reconstruction of tissue engineered corneal epithelium.

Key words: adipose-derived stem cells, toxicity evaluation, green fluorescent protein, transfection, rabbit