1.Genetic modification of bone marrow mesenchymal stem cells with human CXCR4 gene and migration in vitro.
Yue ZHANG ; Lailiang OU ; Zhaokang CHENG ; Xiaohua JIA ; Nianfa GAO ; Deling KONG
Journal of Biomedical Engineering 2009;26(3):595-600
This study was amied to construct CXCR4 gene modified bone marrow mesenchymal stem cells (MSCs), and investigate the effect of CXCR4 expression on MSCs migration. The retrovirus vector pMSCV-CXCR4-IRES-GFP that expresses human CXCR4 gene was cloned,the MSCs were transduced by the virus, and the expression of OXCR4 was analyzed by FACS, RT-PCR and immunofluorescence staining. The migration assay was performed using Transwell method in the presence of SDF-1. FACS results showed that 46% of the transduced MSCs were CXCR4 positive, and 57% were GFP positive. The expression of CXCR4 in MSCs was also confirmed by RT-PCR and immunostaining. The migration of MSCs was induced by SDF-1 and was strongly dependent on CXCR4 expression. The concentration of SDF-1 had effect on the migration and the transmigration rate of CXCR4 modified; the amount of MSCs was 5-fold higher than that of untransduced MSCs when the optimal concentration rose to 50 ng/ml. These data indicate that SDF-1/CXCR4 plays an important role in MSCs migration ,and the CXCR4 genetic modification approach could be applied to enhance cell homing, and engraftment in MSCs therapy.
Bone Marrow Cells
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cytology
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Cell Movement
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Cells, Cultured
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Chemokine CXCL12
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pharmacology
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Genetic Vectors
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genetics
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Humans
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Mesenchymal Stromal Cells
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cytology
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metabolism
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Receptors, CXCR4
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biosynthesis
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genetics
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Retroviridae
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genetics
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metabolism
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Transduction, Genetic
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Transfection