[Genetic modification of bone marrow mesenchymal stem cells with human CXCR4 gene and migration in vitro].
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.
Zhang
Yue
Y
College of Biotechnology, Tianjin University of Science & Technology, Tianjin Key Laboratory of Industrial Microbiology, Tianjin 300457, China.
Ou
Lailiang
L
Cheng
Zhaokang
Z
Jia
Xiaohua
X
Gao
Nianfa
N
Kong
Deling
D
D001854:Bone Marrow Cells / Q000166:cytology; D002465:Cell Movement; D002478:Cells, Cultured; D054377:Chemokine CXCL12 / Q000494:pharmacology; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism*; D019718:Receptors, CXCR4 / Q000096:biosynthesis / Q000235:genetics*; D012190:Retroviridae / Q000235:genetics / Q000378:metabolism; D014161:Transduction, Genetic; D014162:Transfection
C527276:CXCR4 protein, human; D054377:Chemokine CXCL12; D019718:Receptors, CXCR4