中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (23): 4277-4281.doi: 10.3969/j.issn.1673-8225.2011.23.022

• 干细胞与中医药 stem cells and traditional Chinese medicine • 上一篇    下一篇

枸杞多糖对精原干细胞体外增殖的影响

马良宏1，邱志军2，闫圣男2，冯立新3，王燕蓉4，李培军1，陈福宝1   

  1. 1宁夏医科大学附属医院泌尿外科，宁夏回族自治区银川市 750004
    2宁夏医科大学，宁夏回族自治区银川市750004
    3上海交通大学医学院生殖干细胞实验室，上海市 200025
    4宁夏医科大学生育力保持教育部重点实验室，宁夏回族自治区银川市750004
  • 收稿日期:2011-03-22 修回日期:2011-05-15 出版日期:2011-06-04 发布日期:2011-06-04
  • 通讯作者: 陈福宝，教授，宁夏医科大学附属医院泌尿外科，宁夏回族自治区银川市750004
  • 作者简介:马良宏☆，男，1975年生，宁夏回族自治区银川市人，回族，2006年自复旦大学毕业，博士，副主任医师，主要从事男性不育的应用基础研究。 mmm1770@163.com 并列第一作者：邱志军★，男，1983年生，湖南省耒阳市人，汉族，宁夏医科大学在读硕士，医师，主要从事精原干细胞的体外增殖研究。
  • 基金资助:

    国家自然科学基金资助项目(30860282)；宁夏自然科学基金资助项目(NZ08107）。

Influence of lycium barbarum polysaccharides on proliferation of spermatogonial stem cells in vitro

Ma Liang-hong1, Qiu Zhi-jun2, Yan Sheng-nan2, Feng Li-xin3, Wang Yan-rong4, Li Pei-jun1, Chen Fu-bao1   

  1. 1Department of Urology, Affiliated Hospital of Ningxia Medical University, Yinchuan  750004, Ningxia Hui Autonomous Region, China
    2Ningxia Medical University, Yinchuan  750004, Ningxia Hui Autonomous Region, China
    3Laboratory for Germline Stem Cell Research, Shanghai Jiao Tong University School of Medicine, Shanghai  200025, China
    4Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education in Ningxia Medical University, Yinchuan  750004, Ningxia Hui Autonomous Region, China
  • Received:2011-03-22 Revised:2011-05-15 Online:2011-06-04 Published:2011-06-04
  • Contact: Chen Fu-bao, Professor, Department of Urology, Affiliated Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • About author:Ma Liang-hong☆, Doctor, Associate chief physician, Department of Urology, Affiliated Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China mmm1770@163.com Ma Liang-hong and Qiu Zhi-jun contributed equally to this paper.
  • Supported by:

    the National Natural Science Foundation of China, No.30860282*; the Natural Science Foundation of Ningxia, No. NZ08107*

PDF

492

被引次数

28

摘要：

背景：精原干细胞移植对不育具有潜在的临床应用价值，体外建立精原干细胞的培养系统获得数量较多的精原干细胞，仍是目前研究中亟待解决的问题。
目的：观察枸杞多糖对精原干细胞体外增殖的影响。
方法：采用两步酶消化法获取出生4~6 d雄性C57BL/6小鼠睾丸Sertoli细胞与精原干细胞，将精原干细胞接种在Sertoli细胞饲养层上，再加入枸杞多糖或联合细胞因子添加到细胞培养液中。1周后以流式细胞仪检测细胞周期及细胞活性率，并检测各组精原干细胞GFRa-1、Thy-1、c-kit的阳性率。
结果与结论：单独加入枸杞多糖后精原干细胞数量明显增加，增殖明显，联合加入胶质细胞源性神经营养因子与白血病抑制因子精原干细胞增殖更加明显(P < 0.05)。并发现体外培养1周后的精原干细胞仍保持其睾丸组织内的精原干细胞特征，大多仍维持在未分化状态。表明在枸杞多糖或枸杞多糖联合胶质细胞源性神经营养因子及白血病抑制因子作用下，可促进精原干细胞体外增殖。

关键词: 精原干细胞, 枸杞多糖, 体外, 培养, 增殖

Abstract:

BACKGROUND: It has potential clinical value to use spermatogonial stem cell transplantation to cure infertility. In order to obtain a large number of spermatogonial stem cells for transplantation, culture system in vitro need to be established, which is still an urgent problem at present.
OBJECTIVE: To investigate the effect of lycium barbarum polysaccharides (LBP) on proliferation of spermatogonial stem cells in vitro.
METHODS: Sertoli cells and spermatogonial stem cells were separated from testis of 4-6 days postpartum C57BL/6 male mice by two-step enzyme digestion method. The spermatogonial stem cells were seeded on the sertoli cells feeder layer and cocultured in vitro with LBP or cytokines in cell culture medium. After one-week coculture, the cell cycle and cell activity rate of spermatogonial stem cells were determined by flow cytometry. Positive rate of spermatogonial stem cells for GFRa-1, Thy-1, c-kit as cell markers were detected.
RESULTS AND CONCLUSION: Adding LBP into the culture medium alone, the amount of spermatogonial stem cell was increased apparently, and spermatogonial stem cells proliferated significantly. Adding LBP, glial cell line-derived neurotrophic factor and leukemia inhibitory factor into the culture medium together, spermatogonial stem cells proliferated more significantly (P < 0.05). Also spermatogonial stem cells after 1-week in vitro culture maintained the feature of spermatogonial stem cells within the testicular tissue, most of them remained undifferentiated state. Results indicate that LBP or combination of some cytokines can promote the proliferation of spermatogonial stem cells in vitro.

中图分类号: