解剖学报

解剖学报 ›› 2013, Vol. 44 ›› Issue (3 ): 334-338.doi: 10.3969/j.issn.0529-1356.2013.03.008

• 细胞和分子生物学 • 上一篇    下一篇

小鼠睾丸支持细胞对人脐带间充质干细胞增殖能力的影响

张芬熙 洪艳* 梁文妹   

  1. Department of Histology and Embryology of Guiyang Medical College, Guiyang 550004, China
  • 收稿日期:2012-07-09 修回日期:2013-01-11 出版日期:2013-06-06 发布日期:2013-07-16
  • 通讯作者: 洪艳 E-mail:276953488@QQ.com
  • 基金资助:

    贵州省社会发展科技攻关项目 黔科合SY[2009]3071；贵州省教育厅项目，黔教高发[2010]305;省自然科学基金资助项目

Effect of mouse Sertoli cells on human umbilical cord mesenchymal stem cells proliferation

ZHANG Fen-xi  HONG Yan*  LIANG Wen-mei   

  • Received:2012-07-09 Revised:2013-01-11 Online:2013-06-06 Published:2013-07-16

摘要：

目的 探讨睾丸支持细胞（SCs）对体外培养人脐带间充质干细胞(hUCMSCs）增殖能力的影响。方法 体外分离培养并鉴定hUCMSCs和SCs；
Transwell-insert系统共培养SCs 和hUCMSCs（共培养组）；普通DMEM-F12培养基培养的hUCMSCs作为对照（对照组）；添加细胞因子
(白细胞介素-3，粒细胞-巨噬细胞集落刺激因子) 的DMEM-F-12培养基培养的hUCMSCs作为阳性对照 (细胞因子组）。用流式细胞术、免疫细胞
化学等方法检测SCs对hUCMSCs增殖、细胞周期、表面标记分子表达的影响，透射电镜观察细胞超微结构。结果 与对照组相比，共培养组和细胞
因子组hUCMSCs增殖明显加快，以共培养组尤为显著（P<0.05）；共培养组和细胞因子组hUCMSCs 表面分子CD29和CD105阳性率均增高；细胞周
期分析显示，共培养组G0/G1期细胞数与对照组相比无明显改变，而细胞因子组略有减少。透射电镜观察对照组与共培养组细胞均呈现原始细胞
的超微结构特点。结论 SCs共培养可促进hUCMSCs增殖，且共培养后仍保持其干细胞特性。

关键词: 脐带间充质干细胞, 支持细胞, 共培养, 体外培养, 免疫组织化学, 流式细胞术, 小鼠

Abstract:

Objective To observe the regulatory effect of Sertoli cells (SCs) on proliferation in human umbilical cord mesenchymal
stem cells (hUCMSCs) in vitro. Methods hUCMSCs and SCs were isolated, cultured and identified in vitro. A co-culture system was
established by culturing SCs in the Transwell insert and hUCMSCs on the plastic plates(the co-culture group). hUCMSCs culture in
DMEM-F12 medium was used as the control group. hUCMSCs culture in DMEM-F12 medium supplemented IL-3 and GM-CSF served as the
cytokine group. Proliferation, cell cycle analyse and surface marker molecules of hUCMSCs were studied by flow cytometry and
immunocytochemistry. Ultrastructures of the cultured cells were observed by transmission electron microscopy. Results Compared
with the control, proliferation of hUCMSCs in the co-culture- and cytokine-group increased significantly in the co-culture group
(P<0.05). The expression of CD29 and CD105 on the surface of hUCMSCs in the co-culture and the cytokine group also increased.
Cell cycle analysis showed that there were a few cells arrested at quiescent phases in the co-culture and cytokine groups,
especially in the cytokine group. In the coculture group and the control group，hUCMSCs may displayed the ultrastructural features
of the primitive cells by transmission electron microscopy. Conclusion Proliferation of hUCMSCs may be promoted as co-culture with
SCs and still kept their stem cell properties after the co-culture.

Key words: Umbilical cord mesenchymal stem cell, Sertoli cell, Co-culture, Culture in vitro, Immunohistochemistry, Flow cytometry, Mouse

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