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Effect of stromal cell derived factor-1α/CXCR4/CXCR7 axis on migration of the bone marrow mesenchymal stem cells
Pages: 639-645
Year: Issue:  5
Journal: Acta Anatomica Sinica

Keyword:  Stromal cell derived factor-1αBone marrow mesenchymal stem cellWestern blottingRat;
Abstract: Objective To investigate expression of CXCR4 and CXCR7 protein and mRNA, which are the receptors of stromal cell derived factor-1α(SDF-1α), in the bone marrow mesenchymal stem cells (BMSCs);to explore the role of SDF-1α/CXCR4/CXCR7 axis in migration of BMSCs in vitro and the possible mechanism .Method BMSCs were isolated from rats and cultured in vitro.CD29, CD44 and CD34 of the cells were identified by flow cytometry .CXCR4-selective antagonist AMD 3100 and CXCR7-specific neutralizing antibody were applied to block CXCR 4 and CXCR7 respectively.The expressions of CXCR4 and CXCR7 mRNA and protein on BMSCs were detected with RT-PCR and Western blotting .Transwells chamber test was used to observe the migration of BMSCs .The BMSCs were divided into the BMSCs group ( A ) , the AMD3100 pretreated BMSCs group ( B ) , the CXCR7-specific neutralizing antibody pretreated BMSCs group(C), the AMD3100 +CXCR7-specific neutralizing antibody pretreated BMSCs group ( D).Result Flow cytometry showed that the expressions of CD 44 and CD29 were positive, while the expression of CD34 was negative in the third passage of BMSCs (P3-BMSCs).CXCR4 and CXCR7 protein and mRNA were both expressed in P3-BMSCs. Compared with the A group, the expression of CXCR4 and CXCR7 protein declined significantly in the B group and the D group;the protein expression of CXCR7 in the C group was lower compared with the A group (P<0.05).However, the expression of CXCR4 mRNA and CXCR7 mRNA had no significant difference between groups .SDF-1αfactor promoted migration of BMSCs ( P <0.05 ).Compared with the 0μg/L group, the numbers of migrated cells were increased significantly in both of the 10μg/L group and the 100μg/L group ( P<0.01 ) .The number of migration of BMSCs was significantly higher in the 100μg/L group than that of the 10μg/L group ( P <0.01 ) .AMD3100 and CXCR7-specific neutralizing antibody both inhibited significantly the migration of BMSCs ( P<0.05 ) , and the attenuate effect was more significant when they worked together ( P<0.05 ) .Conclusion CXCR4 and CXCR7 receptors are co-expressed in P3-BMSCs;the SDF-1αfactor can promote the migration of BMSCs in the concentration dependent manner ;SDF-1α/CXCR4/CXCR7 axis is involved in the migration of BMSCs , and both of the CXCR4 and CXCR7 receptors have a synergistic promoting effect to the BMSCs migration .
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