不同来源干细胞条件培养基对人脐静脉内皮细胞作用的比较

Comparison of the effects of conditioned medium of stem cells from different sources on hu⁃man umbilical vein endothelial cells

目的:研究比较人脐带间充质干细胞(hUCMSCs)和人牙髓干细胞(hDPSCs)来源条件培养基(CM)对人脐静脉内皮细胞(HUVECs)血管生成功能的影响。方法:实验分为NC组、ECM培养基组、hUCMSCs-CM组和hDPSCs-CM组。通过细胞计数试剂盒(CCK-8)法、细胞迁移实验(Transwell)、划痕、成管实验检测HUVECs在不同条件培养基中增殖、迁移、成管的功能,实时荧光定量聚合酶链式反应(RT-qPCR)检测血管生成相关基因的表达,ELISA检测hUCMSCs-CM和hDPSCs-CM中VEGF、HGF、HIF-1α水平。结果:hUCMSCs-CM组和hDPSCs-CM组均能促进HUVECs的增殖、迁移和成管(P<0.05),但两种条件培养基之间未见明显差异(P>0.05);RT-qPCR结果显示,VEGF、KDR、HGF、Ang-2、HIF-1α在hUCMSCs-CM组和hDPSCs-CM组的表达量高于ECM培养基组和NC组(P<0.05);ELISA结果显示,hUCMSCs-CM组和hDPSCs-CM组的VEGF、HGF和HIF-1α蛋白浓度高于NC组,hUCMSCs-CM组中HGF和HIF-1α水平高于hDPSCs-CM组(P<0.05)。结论:hUCMSCs-CM和hDPSCs-CM均能促进HUVECs增殖、迁移和成管,hUCMSCs和hDPSCs均通过分泌多种血管形成相关因子促进HUVECs的血管形成,hUCMSCs表现出更强的分泌能力。

Objective:To compare the effects of human umbilical cord mesenchymal stem cells(hUCMSCs)and human dental pulp stem cells(hDPSCs)derived conditioned medium(CM)on angiogenesis of human umbilical vein endothelial cells(HUVECs).Methods:The experiment was divided into NC group,ECM medium group,hUCMSCs-CM group and hDPSCs-CM group.The proliferation,migration and tube formation of HUVECs in different conditioned medium were detected by CCK-8 method,Transwell,scratch assay and tube formation assay.The expressions of angiogenesis related genes were detected by real-time reverse transcription quantitative PCR(RT-qPCR).The levels of VEGF,HGF and HIF-1αin hUCMSCs-CM and hDPSCs-CM were detected by ELISA.Results:Both hUCMSCs-CM group and hDPSCs-CM group could promote the proliferation,migration and tube formation of HUVECs(P<0.05),but there was no significant difference between the two conditioned media(P>0.05).The results of RT-qPCR showed that the expressions of VEGF,KDR,HGF,Ang-2 and HIF-αin hUCMSCs-CM group and hDPSCs-CM group were higher than that in ECM medium group and NC group(P<0.05).The results of ELISA showed that the protein concentrations of VEGF,HGF and HIF-1αin hUCMSCs-CM group and hDPSCs-CM group were higher than those in NC group,and the levels of HGF and HIF-1αin hUCMSCs-CM group were higher than those in hDPSCs-CM group(P<0.05).Conclusion:Both hUCMSCs-CM and hDPSCs-CM can promote the proliferation,migration and angiogenesis of HUVECs.Both hUCMSCs and hDPSCs promote the angiogenesis of HUVECs by secreting a variety of angiogenesis-related factors,and hUCMSCs shows a stronger secretory capacity.