人脐带和脂肪来源间充质干细胞培养上清对内皮细胞血管新生作用

Effect of human Wharton′s Jelly or adipose derived mesenchymal stem cells culture supernatant on endothelial cells angiogenesis

目的比较人脐带Wharton′s Jelly来源间充质干细胞(WJ-MSC)与脂肪来源间充质干细胞(AD-MSC)培养上清对内皮细胞血管新生的促进作用。方法酶消化法分离获得WJ-MSC及AD-MSC,分别收集其培养上清并与内皮细胞共培养。实时荧光定量逆转录聚合酶链反应检测内皮细胞中促进血管新生相关基因和抑制血管新生基因的表达;Matrigel体外成管实验检测其对内皮细胞管网状结构形成的作用。结果WJ-MSC和AD-MSC培养上清分别与内皮细胞共培养后,内皮细胞促血管新生相关基因的表达水平显著升高,抑制血管新生相关基因表达降低(P<0.01),与对照组相比,WJ-MSC组及AD-MSC组内皮细胞体外管网状结构的形成均显著高于对照组(43.2 mm±9.2 mm和94.3 mm±13.2 mm,86.1 mm±7.2 mm,P<0.01)。结论WJ-MSC与AD-MSC培养上清对内皮细胞血管新生均具有促进作用,且二者的促进作用相当。

Objective To compare the curative effect of mesenchymal stem cells derived from human Wharton′s Jelly(WJ-MSC)or adipose(AD-MSC)culture supernatant on endothelial cells angiogenesis.Methods WJ-MSC and AD-MSC were isolated,identified,and the culture supernatant of stem cells was collected.The WJ-MSC or AD-MSC supernatant co-cultured with the endothelial cells.The expression levels of pro-angiogenic and anti-angiogenic genes of endothelial cells were assessed using qRT-PCR analysis,and the effects of stem cell culture supernatant on angiogenesis were evaluated by performing a tube formation assay in vitro.Results After adding WJ-MSC and AD-MSC culture supernatant,the expression levels of pro-angiogenic genes in endothelial cells were upregulated,and the expression levels of anti-angiogenic genes were downregulated significantly in both experimental groups compared to the control group(P<0.01),and tube formation of endothelial cells was also significantly increased in both experimental groups as determined by the increase of the tube length((43.2±9.2)mm vs(94.3±13.2)mm,(86.1±7.2)mm,P<0.01).Conclusion The results showed that AD-MSC culture supernatant can promote endothelial cells angiogenesis and its curative effect is similar to that of WJ-MSC.