中国组织工程研究 ›› 2014, Vol. 18 ›› Issue (37): 5955-5960.doi: 10.3969/j.issn.2095-4344.2014.37.009

• 脐带脐血干细胞 umbilical cord blood stem cells • 上一篇    下一篇

人脐血间充质干细胞来源的外泌体:分离鉴定及生物学特性

张  娟1,刘  峰2,张  薇2,丛  旭2,王彩生1,3,魏  来2   

  1. 1内蒙古医科大学研究生学院,内蒙古自治区呼和浩特市  010110
    2北京大学人民医院,北京大学肝病研究所,丙型肝炎和肝病免疫治疗北京市重点实验室,北京市  100044
    3呼和浩特市第二医院,内蒙古自治区呼和浩特市  010031
  • 修回日期:2014-07-31 出版日期:2014-09-03 发布日期:2014-09-03
  • 通讯作者: 王彩生,硕士,主任医师,内蒙古医科大学研究生学院,内蒙古自治区呼和浩特市 010110;呼和浩特市第二医院,内蒙古自治区呼和浩特市 010031 并列通讯作者:魏来,博士,教授,北京大学人民医院,北京大学肝病研究所,丙型肝炎和肝病免疫治疗北京市重点实验室,北京市 100044
  • 作者简介:张娟,女,1988年生,河北省玉田县人,汉族,内蒙古医科大学免疫学在读硕士,医师,主要从事干细胞的基础与临床应用研究。

Exosomes derived from human umbilical cord blood mesenchymal stem cells: isolation, identification and biological characteristics

Zhang Juan1, Liu Feng2, Zhang Wei2, Cong Xu2, Wang Cai-sheng1, 3, Wei Lai2   

  1. 1Faculty of Graduate Studies, Inner Mongolia Medical University, Hohhot 010110, Inner Mongolia Autonomous Region, China
    2Peking University People’s Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, Beijing 100044, China
    3the Second Hospital of Hohhot City, Hohhot 010031, Inner Mongolia Autonomous Region, China
  • Revised:2014-07-31 Online:2014-09-03 Published:2014-09-03
  • Contact: Wang Cai-sheng, Master, Chief physician, Faculty of Graduate Studies, Inner Mongolia Medical University, Hohhot 010110, Inner Mongolia Autonomous Region, China; the Second Hospital of Hohhot City, Hohhot 010031, Inner Mongolia Autonomous Region, China Corresponding author: Wei Lai, M.D., Professor, Peking University People’s Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, Beijing 100044, China
  • About author:Zhang Juan, Studying for master’s degree, Physician, Faculty of Graduate Studies, Inner Mongolia Medical University, Hohhot 010110, Inner Mongolia Autonomous Region, China

摘要:

背景:外泌体是间充质干细胞分泌的膜性小囊泡,越来越多的研究表明,间充质干细胞可能通过旁分泌外泌体而发挥对组织损伤的修复作用,目前人脐血间充质干细胞来源的外泌体分离鉴定尚未见报道。
目的:分离纯化人脐血间充质干细胞来源的外泌体,并鉴定其生物学特性。
方法:收集人脐血间充质干细胞培养上清,采用超滤及梯度离心法分离并纯化外泌体,通过透射电镜观察其形态,流式细胞术检测其表面标志 CD63、CD81、CD90、CD73、CD105、CD29、CD166。
结果与结论:人脐血来源的间充质干细胞能够分泌外泌体,电镜下呈椭圆或碟状的囊泡结构,直径40-100 nm。外泌体表达其共性表达标志CD63、CD81及间充质干细胞表面黏附分子CD90、CD73、CD105、CD29、CD166。结果表明采用超滤及梯度离心法可以成功从间充质干细胞培养上清中分离纯化外泌体,间充质干细胞分泌的外泌体具有外泌体的胞膜蛋白组分。


中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程


全文链接:

关键词: 干细胞, 脐带脐血干细胞, 外泌体, 人脐血间充质干细胞, 分离鉴定, 活性

Abstract:

BACKGROUND: Exosomes are membrane vesicles secreted by mesenchymal stem cells. Increasing studies have shown that mesenchymal stem cells can secrete exosomes via paracrine function to play a role in tissue injury. However, reports on how to isolate and identify exosomes derived from human umbilical cord blood mesenchymal stem cells are few.
OBJECTIVE: To extract, purify and identify exosomes derived from human umbilical cord blood mesenchymal stem cells.
METHODS: The cell culture supernatant of human umbilical cord blood mesenchymal stem cells was collected. Exosome was extracted and purified with ultrafiltration and gradient centrifugation methods. The morphology of exosome was observed by transmission electronic microscope, and the expressions of CD63, CD81, CD90, CD73, CD105, CD29, and CD166 in exosome of mesenchymal stem cells were analyzed by fluorescent activated cell sorting.
RESULTS AND CONCLUSION: Mesenchymal stem cells from human umbilical cord blood secreted exosome 
which exhibited elliptic or saucer-like shape and its diameter ranged from 40 to 100 nm with membrane structure. Exosome could express the common surface adhesion molecules CD63, CD81 and the surface adhesion molecules CD90, CD73, CD105, CD29, CD166 of mesenchymal stem cells. These findings indicate that exosome may be secreted by mesenchymal stem cells of human umbilical cord blood, which contains plasma membrane proteins of umbilical cord blood mesenchymal stem cells.


中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程


全文链接:

Key words: fetal blood, mesenchymal stem cells, exosomes, membrane proteins

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