摘要
背景:人骨骼肌源性血管内皮细胞位于血管壁,共表达肌肉干细胞和血管内皮细胞的标记(CD56^+CD34^+CD144^+CD45^-)。研究显示,人肌血管内皮细胞与间充质干细胞存在相似性,表达间充质干细胞表面标记物,具有多向分化潜能。目的:建立人肌血管内皮细胞作为滋养层与人脐血CD34^+细胞体外培养体系,以培养前后CD34^+细胞数量变化、免疫表型及集落形成能力为指标,评估人肌血管内皮细胞对造血干/祖细胞的体外支持效力。方法:实验分3组:①实验组:以人肌血管内皮细胞为滋养层与人脐血CD34~+细胞体外共培养;②对照组:以人骨髓间充质干细胞为滋养层与人脐血CD34^+细胞体外共培养;③空白对照组:人脐血CD34~+细胞单独培养。共培养第1,2,4周分析人脐血CD34^+细胞数量、血细胞免疫表型和造血干/祖细胞集落形成能力等指标,第5周时因无细胞存活故不进行检测。结果与结论:①人脐血CD34^+细胞数量变化:在1,2,4周时实验组比对照组有所增加,但二者差异均无显著性意义(P>0.05);②流式细胞术分析血细胞免疫表型:只有在第2周时对照组人脐血CD34^+细胞CD34^+CD33^-表达率略高于实验组,差异有显著性意义(P<0.05),其他亚型免疫表型CD45^+、CD14^+、CD10^+/CD19^+表达两组之间比较差异无显著性意义(P>0.05);③造血干/祖细胞集落形成能力:在1,2,4周时实验组与对照组造血干/祖细胞集落数量差异无显著性意义(P>0.05);④由于空白对照组无滋养层,人脐血CD34^+细胞数量在1周时明显下降,第2周计数无存活,因此未进行血细胞免疫表型及集落分析;⑤结果表明,人肌血管内皮细胞作为滋养层细胞具有与人骨髓间充质干细胞相似的体外造血支持作用。
BACKGROUND:Human skeletal muscle derived myoendothelial cells(MECs)are located in the vascular wall and co-express the markers of muscle stem cells and vascular endothelial cells(CD56^+CD34^+CD144^+CD45^-).Studies have shown that MECs are similar to mesenchymal stem cells,express the surface markers of mesenchymal stem cells and have the potential of multidirectional differentiation.OBJECTIVE:To establish an in vitro culture system for human umbilical cord blood CD34^+cells with MECs as trophoblastic layer,and to evaluate the in vitro supporting effect of human skeletal muscle MECs on hematopoietic stem/progenitor cells by measuring the changes in the number,immunophenotype and colony forming ability of CD34^+cells before and after culture.METHODS:There were three groups in the experiment.In experimental group,human umbilical cord blood CD34^+cells were co-cultured with MECs as the nourishing layer;in control group,human umbilical cord blood CD34^+cells were co-cultured with bone marrow mesenchymal stem cells as the nourishing layer;and in blank control group,human umbilical cord blood CD34^+cells were cultured alone without the nourishing layer.The main outcome measures,including the number of human umbilical cord blood CD34^+cells,immunophenotype of blood cells and colony formation ability of hematopoietic stem/progenitor cells were analyzed and compared at 1,2,and 4 weeks after co-culture.No detection was conducted at 5 weeks due to the lack of survived cells.RESULTS AND CONCLUSION:(1)The number of human umbilical cord blood CD34+cells increased by MECs as the nourishing layer compared with bone marrow mesenchymal stem cells as the nourishing layer at 1,2,and 4 weeks;however,there was no significant difference between the two groups(P>0.05).(2)The cell immunophenotype by flow cytometry analysis indicated that only in the 2nd week,the expression of CD34^+CD33^-in human umbilical cord blood CD34^+cells in the control group was significantly higher in the experimental group(P<0.05).Other subtypes of immunophenoty
作者
古晶晶
周芮
杨婷婷
杨小萍
许飞
郑波
Gu Jingjing;Zhou Rui;Yang Tingting;Yang Xiaoping;Xu Fei;Zheng Bo(Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;the People’s Hospital of Jiaozuo City,Jiaozuo 454000,Henan Province,China;Human Stem Cell Institute,General Hospital of Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Department of Hematology,General Hospital of Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China)
出处
《中国组织工程研究》
CAS
北大核心
2021年第1期50-55,共6页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金项目(81560023),项目负责人:郑波
2015年度宁夏留学人员科技活动项目,项目负责人:郑波。
关键词
人骨骼肌源性血管内皮细胞
脐血CD34+细胞
体外支持
共培养
滋养层
human skeletal muscle derived myoendothelial cells
umbilical cord blood CD34+stem cells
supporting in vitro
co-culture
nourishing layer
