The Experiment of Co-culture Mesenchymal Stem Cells with Umbilical Vein Endothelial Cells under Hypo

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  Objective By co-culture of hMSCs and HUVECs in hypoxia,simulate a microenvironment similar to where transplantation of hMSCs for treatment of ANFH and investigate the effect of human mesenchymal stem cells (hMSCs) on survival,apoptosis,migration and angiogenesis of human umbilical vein endothelial cells (HUVECs) under the hypoxic condition in vitro.Methods hMSCs and HUVECs were cultured and identified in vitro.Three kinds of conditioned medium,CdM-CdMNOR,CdM-CdMHYP and HUVEC-CdMHYP were prepared.HUVECs were cultured with the above-mentioned conditioned medium in hypoxia.The survival rate,apoptosis rate,migration and angiogenesis of HUVECs was respectively detected by CCK-8,flow cytometry,Tran swell and tube formation assay.The content of SDF-1α,VEGF and IL-6 in CdM was assayed by ELISA.Result hMSCs and HUVECs were cultured and identified successfully.Compared with MSC-CdMNOR and HUVEC-CdMHYP group,the survival rate,migration and angiogenesis of HUVECs in MSC-CdMHYP group was obviously increased while the apoptosis rate was declined (p < 0.05).Moreover,the expression of SDF-1α,VEGF and IL-6 in MSC-CdMYP group was up-regulated.Conclusion In hypoxia,the apoptosis of HUVECs can be inhibited while survival,migration and angiogenesis can be improved by co-culture of hMSCs and HUVECs.The mechanism may be that hMSCs could secrete several cytokines and improve niche to treatment femoral head necrosis.
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