大鼠骨髓间充质干细胞来源微颗粒的提取及鉴定  

Isolation and identification of rat bone marrow mesenchymal stem cells-derived microparticles

作　　者：马瑞君 王苗苗 封启龙[1] MA Rui-jun;WANG Miao-miao;FENG Qi-long(Key Laboratory of Cellular Physiology,Ministry of Education,Shanxi Medical University,Taiyuan 030001,Shanxi Province,China)

机构地区：[1]山西医科大学生理学系细胞生理学教育部重点实验室,山西太原030001 [2]山西医科大学第二附属医院心内科,山西太原030001

出　　处：《中国生物制品学杂志》2022年第2期215-219,共5页Chinese Journal of Biologicals

基　　金：山西省自然科学基金(201901D111195);山西省归国留学人员基金资助项目(2017-055);山西省“1331工程”重点学科建设计划经费(XK201708);山西省重点实验室建设项目(2015012003-08).

摘　　要：目的分离并纯化SD大鼠骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs),在此基础上收集骨髓间充质干细胞来源的微颗粒(bone mesenchymal stem cell-derived microparticles,BMSC-MPs),并对其进行鉴定。方法采用全骨髓贴壁法分离纯化BMSCs,显微镜下观察BMSCs的形态,绘制第5代BMSCs的生长曲线,通过流式细胞术鉴定BMSCs的表面标记。通过血清饥饿法诱导BMSCs释放BMSC-MPs,流式细胞术鉴定BMSC-MPs的数量和表面标记。结果分离的BMSC生长状态良好,呈梭形和典型的“鱼群样”分布,表面标记CD90和CD29阳性率分别为(99.81±0.94)%和(99.90±0.66)%,CD45和CD34阳性率分别为(0.89±0.31)%和(1.79±0.33)%;流式细胞术鉴定BMSC-MPs数量为(2.55±0.86)×10^(7)个/mL,BMSC-MPs表面标记CD90和CD29阳性率分别为(97.46±1.85)%和(94.00±2.03)%,CD45和CD34阳性率分别为(4.89±1.05)%和(1.81±0.33)%。结论成功分离了大鼠BMSCs,在此基础上制备了BMSC-MPs,并进行了初步鉴定,为后续研究BMSC-MPs的功能奠定了基础。Objective To isolate and purify the bone marrow mesenchymal stem cells(BMSCs)of SD rats,and collect and identify the BMSCs-derived microparticles(BMSC-MPs).Methods BMSCs were isolated and purified by whole bone marrow adherent method,and observed for morphology by microscopy.The growth curve of BMSCs of passage 5 was plotted,while the surface markers were identified by flow cytometry.The release of BMSC-MPs from BMSCs was induced by serum starvation,and the number of surface markers of BMSC-MPs were identified by flow cytometry.Results The isolated BMSCs grew well and showed a fusiform and typical“fish-like”distribution,of which the positive rates of surface markers CD90 and CD29 were(99.81±0.94)%and(99.90±0.66)%,while those of CD45 and CD34 were(0.89±0.31)%and(1.79±0.33)%,respectively.The number of BMSC-MPs determined by flow cytometry was(2.55±0.86)×10^(7)cells/mL,while the positive rates of surface markers CD90 and CD29 were(97.46±1.85)%and(94.00±2.03)%,and those of CD45 and CD34 were(4.89±1.05)%and(1.81±0.33)%,respectively.Conclusion Rat BMSCs were successfully isolated,based on which BMSC-MPs were isolated and preliminarily identified.It laid a foundation of further study on the function of BMSC-MPs.

关 键 词：骨髓间充质干细胞 微颗粒 全骨髓贴壁法 流式细胞术 

分 类 号：Q952.5[生物学—动物学]