摘要
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.
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2012年第3期717-721,共5页
Journal of Experimental Hematology
基金
山东省科技攻关计划(编号2009GG10002027)
山东省自然科学基金(编号2006BS03038&2006BS03019)
关键词
脐带间充质干细胞
骨髓间充质干细胞
内皮细胞
内皮祖细胞
血管内皮生长因子
umbilical cord mesenchymal stem cell
bone marrow mesenchymal stem cell
endothelial cell
endothelial progenitor cell
vascular endothelial growth factor