Comparison of lentiviral vector and adenoviral vector mediated gene transfer into rabbit bone marrow mesenchymal stem cells
10.3969/j.issn.2095-4344.2017.09.006
- VernacularTitle:基因重组腺病毒载体与慢病毒载体转染兔骨髓间充质干细胞的对比
- Author:
Shipeng LI
;
Qiang LI
;
Zhengsong SHI
;
Weiliang CAI
;
Yinkuan DING
;
Xuan TAO
- From:
Chinese Journal of Tissue Engineering Research
2017;21(9):1340-1345
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Both lentiviral vector and adenoviral vector are considered as good vectors for gene mediation, and their differences in transferring bone morphogenetic protein 2 (BMP-2) into rabbit bone marrow mesenchymal stem cells (BMSCs) are unclear. OBJECTIVE: To compare the duration, efficiency and the deviation of exogenous gene expression after rabbit BMSCs transfection using lentiviral vector and adenoviral vector which are used to mediate enhanced green fluorescent proteins (EGFP) and BMP-2. METHODS: Rabbit BMSCs at passage 5 were exposed to Ad-EGFP-BMP-2 (group A) or Lenti-EGFP-BMP-2 (group B) with multiplicity of infection of 100, as transfection groups. And in control group (group C), the same quality of culture medium was required equivalent to the groups A and B. The expression of EGFP was observed by inverted fluorescence microscope at various time intervals. And the expression of exogenous gene BMP 2 in cells was detected and analyzed by immunohistochemical staining at 72 hours after transfection as well as by western blot at 72 hours, 1, 3 weeks after transfection. RESULTS AND CONCLUSION: The intense green fluorescence emerged under the microscope at 24-48 hours after transfection in group A, which was stronger than group B, reached the peak at 72 hours, and then decreased at 1 week until disappearance at 3 weeks. No EGFP expression was detected in group C. High expression of BMP-2 was found in group A but was dramatically downregulated after 1 week. Group B showed the high expression of EGFP/BMP-2 persisted for a longer period after transfected that even lasted for 3 weeks. Overall, the lentiviral vector and adenoviral vector can efficiently transfect rabbit BMSCs and stably express the target gene of EGFP/BMP-2. Under the same MOI, compared to the adenoviral vector, transfection of lentiviral vector to rabbit BMSCs is more effectively and expression of EGFP/BMP-2 can be persistent in a longer term.
