Transfection of rabbit conjunctival epithelial cells with adeno-associated virus mediated enhanced green fluorescent protein
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Supported by National Natural Science Foundation of China (30570954).

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    Abstract:

    Objective:To transfect rabbit conjunctival epithelial cells in vitro with recombinant adeno-associated virus 2-mediated enhanced green fluorescent protein (rAAV2-EGFP),so as to lay a foundation for studying the proliferation and differentiation of conjunctival epithelial stem cells.Methods: Rabbit conjunctival epithelial cells were cultured in vitro.The second passage of rabbit conjunctival epithelial cells were transfected with rAAV2-EGFP in various multiplicity of infection (MOI=104,105,106).The expression of green fluorescent protein was examined 1,3,5,and 7 days after transfection and the transfection rate was calculated.MTT assay was used to study the influence of rAAV2-EGFP on the growth of cells.Results: With the increase of MOI value and the prolongation of transfection,the expression of EGFP was gradually increased and reached its peak on 7-8 days after transfection.MTT assay showed that there was no significant difference between the MOI group and the control group.Conclusion: rAAV2-EGFP can efficiently transfect rabbit conjunctival epithelial cells and have no influence on the proliferation of the cells.

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History
  • Received:February 22,2008
  • Revised:November 07,2008
  • Adopted:December 08,2008
  • Online: December 17,2008
  • Published:
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