Effect of lentiviral vector-mediated HCV receptor OCLN / CD 81 gene transfection on the tree shrew bone marrow mesenchymal stem cells
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(1. the Center of Tree Shrew Germplasm Resources,Institute of Medical Biology,Chinese Academy of Medical Science and Peking Union Medical College; 2. Yunnan Science and Technology Talent and Platform Program,Kunming 650118,China)

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Q95-33

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    Abstract:

    Objective To observe the effect of lentiviral vector-mediated HCV receptor OCLN / CD 81 gene transfection on the biological characteristics and gene expression of tree shrew bone marrow mesenchymal stem cells (BM-MSCs) under in vitro culture conditions. Methods BM-MSCs were obtained by our previously described isolation method.We constructed a lentivirus vector containing the human OCLN / CD 81 gene and transfected it into BM-MSCs. The transfection efficiency was determined by fluorescence inverted microscopy and flow cytometry, and the proliferation of BMMSCs was determined using CCK8. Multipotent differentiation was induced by osteoblasts and adipocytes. The gene and protein expression of OCLN / CD 81 was detected by RT-PCR and WB, respectively. Results The transfection efficiency levels of LV- CD 81 and LV- OCLN were 99. 4% and 22. 6%, respectively, on MOI 100. The cell proliferation trends were similar to those of the untransfected BM-MSCs. The BM-MSCs were still undergoing osteogenic and adipogenic differentiation, but this declined. The pluripotency marker of NANOG mRNA was increased ( P < 0. 05), while the level of LIN28A mRNA was decreased ( P < 0. 05). BM-MSCs could express the transferred gene efficiently after transfection.Conclusions The constructed lentivirus vector containing the human OCLN / CD 81 gene can be successfully transfected into tree shrew BM-MSCs and the target gene is highly expressed. The multipotent differentiation potential of tree shrew BM-MSCs is clearly affected after transfection.

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History
  • Received:September 27,2018
  • Revised:
  • Adopted:
  • Online: January 03,2019
  • Published: