The Effect Of VEGF Modified Adipose Derived Stem Cells Conditioned Medium On Fibroblasts And Endothelial Cells In Vitro

Background: With the increasing prevalence of chronic wounds, it has become a serious social and economic problem, human adipose derived stem cells(HADSCs) can secrete a variety of growth factors and can be used for wound healing, we are trying to through lentivirus vector carrying vascular endothelial growth factor(VEGF) gene transfect HADSCs, acquire the stable secretion of VEGF.Objective: To provide new method of treating wounds, we using lentivirus carrying VEGF gene modify HADSCs, observe whether the VEGF and b FGF are overexpression stably, detecting the effectsof HADSCs-CM on human dermal fibroblast(HDFs) and human umbilical vein endothelial cells(HUVECs) proliferation, migration in vitro.Methods: We isolated the HADSCs by collagenase digestion method, using flow cytometry analysis surface markers and induced the cells into lipoblast, osteoblast, chondroblast. Modified the HADSCs with lentivirus carrying VEGF gene and examined the secretion of VEGF and b FGF in HADSCs-CM, observe the effects of conditioned medium on the proliferation and migration ability of HDFs and HUVECs by CCK-8 assay and scratch wound model.Results: The expression of VEGF and b FGF were significantly promoted in HADSCs-CM after transfected with lentivirus carrying VEGF gene compared to control groups(P<0.05). When the complete medium mixed with VEGF-HADSCs-CM in the proportion of 1:2, it was the most effective conditioned medium on proliferation of cells, and this mixture significantly enhanced cells migration activities too.Conclusion: The VEGF gene can transfected HADSCs successfully, the VEGF and b FGF secretion enhanced through proteins interaction, the VEGF-HADSCs-CM promoted the proliferation and migration abilities of HDFs and HUVECs, suggesting the potential of using VEGF-HADSCs-CM as a therapy method for chronic wounds.