Recombinant adeno-associated virus type 9 transfection of mouse bone marrow mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.2014.50.003

Abstract

Abstract:

BACKGROUND: Recombinant adeno-associated virus type 9 (rAAV9) has a good affinity to myocardialcytes, and it is an ideal carrier for gene therapy of myocardial infarction.
OBJECTIVE: To investigate the transfection efficiency of rAAV9-enhanced green fluorescence protein (eGFP) in transfection of mouse bone marrow mesenchymal stem cells.
METHODS: rAAV9-eGFP was transfected into mouse bone marrow mesenchymal stem cells which were subcultured to the fourth generation at different multiplicities of infection (MOI=1×105, 1×106, 1×107). To find the best condition of transfection, we observed the eGFP expression under the inverted fluorescence microscope at 1-7 days after transfection. Then the transfection efficiency of rAAV9-eGFP under the optimal MOI was determined by flow cytometry.
RESULTS AND CONCLUSION: The cells with rAAV9-eGFP transfection at MOl of 1×107 began to exhibit eGFP expression at the 1st day after transfection, and the cells transfected at MOI of 1×105 and 1×106 began to exhibit eGFP expression from the 2nd day after transfection. The eGFP expression increased with the MOI used for transfection, and the intensity of eGFP expression increased with the time going on. At the 5th day, the eGFP
expression reached the peak value, and meanwhile, the transfection efficiency at MOI=1×107 was about 8%. rAAV9-eGFP cannot transfect mouse bone marrow mesenchymal stem cells effectively.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: mice, bone marrow, mesenchymal stem cells, transfection

CLC Number:

R394.2

Dong Shuo, Huang Ying, Ma Yi-tong, Meng Lin-lin, Liu Fen, Chen Bang-dang, Chen Xiao-cui. Recombinant adeno-associated virus type 9 transfection of mouse bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(50): 8048-8053.

Figures/Tables 5

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