ABSTRACT
Objective To investigate the effects of VEGF-C,SDF-1,MCP-1 and G-CSF on migration and mobilization of CD34~+/CD133~+/VEGFR-3~+ lymphatic endothelial progenitor cells(LEPCs).Methods Mononuclear cells were isolated from canine peripheral blood by non-continuous density centrifugation with Percoll solution.VEGFR-3~+ cells were sorted with flow cytometxy.Expression of specific marks CD34 and CD133 on VEGFR-3~+ cells were observed under a confocal laser scanning microscope.The effects of VEGF-C,SDF-1,MCP-1 and G-CSF on chemotatic migration of LEPCs were examined by transmigration assay.The morphological characteristics of the transmigrated cells were observed under scanning electron microscope.VEGF-C,SDF-1,MCP-1 and G-CSF were administrated to rats by subcutaneous injection.The number of LEPCs in peripheral blood was analyzed with flow cytometry and white blood cells were counted with blood cell analyzer.Results The percentage of the migrated cells through the membrane was greater in the VEGF-C,SDF-1,MCP-1 and G-CSF groups than that in the control group.After the cells were treated with blocking antibodies of VEGFR-3 and CXCR-4,the percentage of the migrated cells decreased obviously.After the injections of VEGF-C,SDF-1,MCP-1 and G-CSF,LEPCs in peripheral blood of rats increased obviously,while significant increase of white blood cells was observed.Conclusion VEGF-C,SDF-1,MCP-1 and G-CSF play a role in the chemotatic migration and mobilization of LEPCs.The signaling pathways of VEGF-C/VEGFR-3 and SDF-1/CXCR-4 have important regulating effects on the chemotatic migration and mobilization of LEPCs.