血管内皮生长因子对人骨髓和脐带间充质干细胞中内皮细胞组分的影响

Influence of Vascular Endothelial Growth Factor on Endothelial Components in Human Bone Marrrow and Umbilical Cord Mesenchymal Stem Cells

本研究旨在比较人骨髓间充质干细胞(BM-MSC)和脐带间充质干细胞(UC-MSC)中的内皮细胞(EC)比例,并探讨血管内皮生长因子(VEGF)对MSC中EC比例的影响。利用流式细胞术检测EC标志CD34+CD133+和vWF+CD31+双阳性细胞的比例,用瑞氏染色观察经VEGF作用的MSC细胞形态变化,免疫荧光技术观察CD34、CD133、CD31、VWF的表达,最后采用实时荧光定量PCR检测VEGF对EC标志性基因表达的影响。结果表明:BM-MSC和UC-MSC中存在少量EC及内皮祖细胞(EPC),经VEGF 10 ng/ml作用24 h后,MSC的长宽比变大,EC比例上升而EPC比例下降,EC相关标志基因Tie-2和ecNOS等的表达均上调,其中UC-MSC对VEGF的反应更明显。结论:BM-MSC和UC-MSC中存在少量的EC及EPC,VEGF可以提高EC的比例和细胞功能。

This study was aimed to compare the proportion of endothelial cells（EC） in human bone mrrrow mesenchymal stem cell（BM-MSC） and human umbilical cord mesenchymal stem cells（UC-MSC）,and to investigate the influence of vascular endothelial growth factor（VEGF） on proportion of EC in MSC.Flow cytometry was used to detect the proportion of CD34＋CD133＋ and vWF＋CD31＋ double positive cells in MSC.Wright’s staining was employed to observe the influence of VEGF on morphology of MSC.The expressions of CD34,CD133,CD31,vWF were detected by immunofluorescence.qRT-PCR was performed to detect the influence of VEGF on EC marker genes’ expression of MSC.The results showed that there were a small amount of EC and endothelial progenitor cells（EPC） in obtained BM-MSC and UC-MSC.After exposed to VEGF 10 ng/ml for 24 h,aspect ratio of MSC and the propotion of EC increased,while proportion of EPC decreased.Expression of EC related marker genes such as Tie-2 and ecNOS up-regulated,especially in UC-MSC.It is concluded that small amount of EC and EPC exists in cultured BM-MSC and UC-MSC,VEGF can enhance the proportion and function of EC in MSC.