[Comparison of transduction efficiencies of various gene vectors in human bone-marrow-derived mesenchymal stem cells].
Zheng Shan Liu, Cheng Zhang, Yan Chang Shang, Fu Xiong, Shan Wei Feng, Yong Li, Yong Xu, Chang Zhou
Abstract
Zheng Shan Liu, Cheng Zhang, Yan Chang Shang, Fu Xiong, Shan Wei Feng, Yong Li, Yong Xu, Chang Zhou
Abstract
OBJECTIVE: To compare the transduction efficiencies of adenoviral vector, adeno-associated viral vector, baculoviral vector, and plasmid vector in human bone-marrow-derived mesenchymal stem cells (hBMSCs). METHODS: The hBMSCs were cultured in vitro and transducted with the adenoviral vector, adeno-associated viral vector, baculoviral vector, and plasmid vector. The expression of target protein was observed by inverted fluorescent microscopy and flow cytometry. RESULTS: Inverted fluorescent microscopy showed that some of the hBMSCs after transduction expressed the green fluorescent protein (GFP) and the hBMSCs transducted with baculoviral vector expressed more GFP than those of other three vectors. Flow cytometry showed that the transduction efficiencies and mean fluorescence intensities of the adenoviral vector, adeno-associated viral vector, and plasmid vector were 42%, 37%, and 22% and 158, 115, and 77, respectively, which were significantly lower than those of baculoviral vector (70%, P < 0.01; 212, P < 0.05; respectively). CONCLUSION: Compared with the adenoviral vector, adeno-associated viral vector, and plasmid vector, the baculoviral vector has higher transduction efficiency in hBMSCs and therefore may be a more suitable gene vector for research in human gene therapy.
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OBJECTIVE: To compare the transduction efficiencies of adenoviral vector, adeno-associated viral vector, baculoviral vector, and plasmid vector in human bone-marrow-derived mesenchymal stem cells (hBMSCs). METHODS: The hBMSCs were cultured in vitro and transducted with the adenoviral vector, adeno-associated viral vector, baculoviral vector, and plasmid vector. The expression of target protein was observed by inverted fluorescent microscopy and flow cytometry. RESULTS: Inverted fluorescent microscopy showed that some of the hBMSCs after transduction expressed the green fluorescent protein (GFP) and the hBMSCs transducted with baculoviral vector expressed more GFP than those of other three vectors. Flow cytometry showed that the transduction efficiencies and mean fluorescence intensities of the adenoviral vector, adeno-associated viral vector, and plasmid vector were 42%, 37%, and 22% and 158, 115, and 77, respectively, which were significantly lower than those of baculoviral vector (70%, P < 0.01; 212, P < 0.05; respectively). CONCLUSION: Compared with the adenoviral vector, adeno-associated viral vector, and plasmid vector, the baculoviral vector has higher transduction efficiency in hBMSCs and therefore may be a more suitable gene vector for research in human gene therapy.
Key concepts: Transduction (biophysics), Viral vector, Green fluorescent protein, Plasmid, Molecular biology, Flow cytometry, Biology, Genetic enhancement