解剖学报

解剖学报 ›› 2019, Vol. 50 ›› Issue (5): 589-594.doi: 10.16098/j.issn.0529-1356.2019.05.008

• 细胞和分子生物学 • 上一篇    下一篇

不同来源间充质干细胞表面标记的比较

杜志朋1 殷国田2 李苗苗1 郭志坤1 李琼1,3*   

  1. 1.新乡医学院河南省医用组织再生重点实验室; 2.新乡医学院第三附属医院心血管内科; 3.新乡医学院护理学院，河南 新乡 453003
  • 收稿日期:2018-07-24 修回日期:2018-09-29 出版日期:2019-10-06 发布日期:2019-12-10
  • 通讯作者: 李琼 E-mail:13513735887@163.com
  • 基金资助:
    CD73通过Nanog促进脂肪间充质干细胞心肌分化和梗死心肌修复的功能与机制研究;脂肪间充质干细胞治疗心肌梗死机制研究;CD73+ADMSC通过CD73/腺苷轴促进梗死心肌修复的免疫调控机制研究

Comparison of surface markers of mesenchymal stem cells from different sources

DU Zhi-peng1 YIN Guo-tian2 LI Miao-miao1 GUO Zhi-kun1 LI Qiong 1,3*   

  1. 1.He’nan Key Laboratory of Medical Tissue Regeneration; 2.Cardiovascular Department, the Third Affiliated Hospital of Xinxiang Medical University； 3.College of Nursing of Xinxiang Medical University, He’nan Xinxiang 453003, China
  • Received:2018-07-24 Revised:2018-09-29 Online:2019-10-06 Published:2019-12-10
  • Contact: LI Qiong E-mail:13513735887@163.com

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摘要：

目的 探讨人脐带间充质干细胞（HUCMSCs）、人脂肪间充质干细胞（ADSCs）和人经血源子宫内膜间充质干细胞（MenSCs）之间表面标记的差异及随培养代次增加表面标记的变化。 方法 分别将HUCMSCs、 ADSCs和MenSCs培养至第3、6、9、12代，用流式细胞术、免疫荧光法检测CD29、CD44、CD73、CD90、CD105、CD45的表达并进行比较。 结果 培养MenSCs和HUCMSCs呈纺锤状，ADSCs形态多样以梭型、多角形为主。流式细胞术检测显示，人HUCMSCs、ADSCs和MenSCs的第3代CD29、CD44、CD73和CD105阳性表达率均在95%以上，CD45阴性表达；第3代MenSCs CD90的表达率为(72.43±0.76)%，均低于其在HUCMSCs（99.67±0.12）%和ADSCs（99.70±0.15）%表达（P＜0.001）。HUCMSCs、ADSCs和MenSCs随培养代次的增加表面标记CD29、CD44、CD73、CD90、CD105和CD45表达率差异无显著性（P>0.05）。免疫荧光结果与流式细胞术结果一致。 结论 HUCMSCs、ADSCs和MenSCs随培养时间的增加稳定高表达 CD29、CD44、CD73、CD90和CD105，不表达CD45，MenSCs的CD90表达率低于其在HUCMSCs和ADSCs的表达。

关键词: 人脐带间充质干细胞, 人脂肪间充质干细胞, 人经血源子宫内膜间充质干细胞, 免疫荧光法, 流式细胞术, 人

Abstract:

Objective To investigate the expression difference of surface markers of human umbilical cord mesenchymal stem cells (HUCMSCs), human adipose mesenchymal stem cells (ADSCs) and human fetal blood source endometrial mesenchymal stem cells (MenSCs) and the changes of surface markers with the culture generation. Methods HUCMSCs, ADSCs and MenSCs were cultured to passage 3, 6, 9 and 12. Five MSC-specific markers, CD29, CD44, CD73, CD90 and CD105, and one HSC markers, CD45, were assessed by flow cytometry and immunofluorescence. Results The cultured MenSCs and HUCMSCs were spindle-shaped, and the ADSCs forms were mainly spindle-shaped and multiple-angular. The results of flow cytometry showed that the MSC-positive markers including CD29, CD44, CD73 and CD105 were highly expressed by at least 95% in the passage 3 HUCMSCs, ADSCs and MenSCs cells and CD45 negative expression. Specifically, CD90 levels of MenSCs of passage 3 was (72.43±0.76)%, which was lower than that in HUCMSCs (99.67±0.12)% and ADSCs (99.70±0.15)%(P<0.001). There was no significant difference in the expression rates of surface markers CD29, CD44, CD73, CD90, CD105 and CD45 with the increase of culture passages in HUCMSCs, ADSCs and MenSCs (P>0.05). The results of immunofluorescence was consistent with those of flow cytometry.Conclusion With the increase of culture time, HUCMSCs, ADSCs and MenSCs are stable in high expression of CD29, CD44, CD73, CD90 and CD105, which does not express CD45, while the CD90 expression rate of MenSCs is lower than that in HUCMSCs and ADSCs.

Key words: Umbilical cord mesenchymal stem cell, Adipose-derived mesenchymal stem cell, Menstrual endometrial mesenchymal stem cell, Immunofluorescence, Flow cytometry, Human