Effects of transfection of CX3CR1 gene on migration of mouse bone marrow-derived mesenchymal stem cells by fractlkine in vitro

Abstract:

Objective：To investigate the roles of Fractalkine in triggering the migration of the mesenchymal stem cells（MSCs） in the Transwell co-culture system. Methods：MSCs were isolated and cultured through the whole bone marrow method and were amplified by the different-speed adherent method. The morphology and biological characteristics of MSCs were examined by flow cytometry. And then the recombinant adenovirus containing CX3CR1 and green fluorescent protein（GFP）（LV-CX3CR1-EGFP） or empty vector（LV-CON-EGFP） were transfected into MSCs. The expression of CX3CR1 mRNA in MSCs was examined by RT-PCR and the pro－tein expression was detected by Western blot in LV-CX3CR1-EGFP infection group，LV-CON-EGFP infection group and non-infec－tion group. The migration of MSCs was detected with Transwell co-culture system. Results：Obtained MSCs were homogeneous in cell morphology with good living condition and the cells showed positive with CD29，CD44，CD34 .There was no expression of CX3CR1 in LV-CON-EGFP group and non-infection group but CX3CR1 can be expressed in LV-CX3CR1-EGFP group. Results of Transwell assays suggested that the migration level of MCSs was higher in LV-CX3CR1-EGFP group than in other groups（P=0.000）. The migrated MSCs were 13.80±2.17/HP，35.80±3.34/HP and 14.80±1.92/HP in LV-CON-EGFP group，LV-CX3CR1-EGFP group and non-infection group respectively. Conclusion：The transfection of CX3CR1 gene could improve the level of migration of MSCs by FKN in vitro.1