中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (2): 282-286.doi: 10.3969/j.issn.1673-8225.2012.02.021

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

小鼠卵巢颗粒细胞适宜培养体系及激素干预的影响****

陈毅斐，许程洁，王鑫泉，徐  营   

  1. 嘉兴学院医学院，浙江省嘉兴市   314001
  • 收稿日期:2011-04-18 修回日期:2011-06-20 出版日期:2012-01-08 发布日期:2012-01-08
  • 通讯作者: 徐营，博士，嘉兴学院医学院，浙江省嘉兴市 314001 Xuyingmrd@yahoo.com
  • 作者简介:陈毅斐，男，1988年生，嘉兴学院医学院临床医学专业本科在读。
  • 基金资助:

    浙江省自然科学基金(Y2110873)、嘉兴市科技局科研重点项目(2007A22013)、嘉兴市科技局科研项目(2010AY1098)、浙江省大学生科技创新活动计划项目(2010R417004)。

Suitable culture system of mouse ovarian granulosa cells and effect of hormone intervention

Chen Yi-fei, Xu Cheng-jie, Wang Xin-quan, Xu Ying   

  1. Jiaxing University College of Medicine, Jiaxing  314001, Zhejiang Province, China
  • Received:2011-04-18 Revised:2011-06-20 Online:2012-01-08 Published:2012-01-08
  • Contact: Xu Ying, Doctor, Jiaxing University College of Medicine, Jiaxing 314001, Zhejiang Province, China Xuyingmrd@yahoo.com
  • About author:Chen Yi-fei, Jiaxing University College of Medicine, Jiaxing 314001, Zhejiang Province, China
  • Supported by:

    the Natural Science Foundation of Zhejiang Province, No. Y2110873*; the Key Programme for Scientific Research of Jiaxing Committee of Science and Technology, No. 2007A22013*; the Programme for Scientific Research of Jiaxing Committee of Science and Technology, No.  2010AY1098*; the College Students' Scientific Research Innovation Projects of Zhejiang Province, No. 2010R417004*

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摘要：

背景：建立稳定、高效的颗粒细胞体外培养体系，可了解颗粒细胞体外生长的特点与基本生物学特性。
目的：建立并优化小鼠卵巢颗粒细胞原代培养体系，探讨孕马血清促性腺激素、人绒毛膜促性腺激素对细胞增殖与细胞骨架的影响。
方法：分离小鼠卵巢颗粒细胞，分别培养于RPMI1640、HTF与BIO-AMF2培养系统，将分离得到的第3代颗粒细胞培养体系中添加人绒毛膜促性腺激素与孕马血清促性腺激素进行干预。
结果与结论：BIO-AMF2培养体系优于RPMI1640、HTF培养体系，原代培养第4天可见细胞汇合成片呈铺路石样生长，颗粒细胞C-kit免疫组化染色呈阳性，细胞周期测定表明体外培养的颗粒细胞大部分处于G1期。在颗粒细胞培养后第2天，激素对颗粒细胞的增殖即显现促进作用，在第4天最为显著，但未发现激素对颗粒细胞的细胞形态与细胞骨架有显著影响。
关键词：卵巢；颗粒细胞；体外培养；激素；细胞骨架
doi:10.3969/j.issn.1673-8225.2012.02.021

关键词: 卵巢, 颗粒细胞, 体外培养, 激素, 细胞骨架

Abstract:

BACKGROUND: The in vitro growth characteristics and basic biological characteristics of granulosa cells can be understood by building a stable and high-efficiency in vitro culture system of granulosa cells.
OBJECTIVE: To build and optimize a primary culture system of mice ovarian granulosa cells, and to explore the effects of pregnant mare serum gonadotropin, human chorionic gonadotropin on the cell proliferation and cytoskeleton.
METHODS: Mouse granulosa cells were separated and cultured in the culture systems of RPMI1640, HTF and BIO-AMF2, respectively. Pregnant mare serum gonadotropin and human chorionic gonadotropin were added to the culture systems of granulose cells in generation 3.
RESULTS AND CONCLUSION: BIO-AMF2 culture system was better than RPMI1640 and HTF culture systems. Cells converged together and represented a cobblestone-like appearance on the 4th day after primary culture. The granulosa cells showed a positive reaction to C-kit immunohistochemical staining. According to cell cycle determination, most of the in vitro cultured granulosa cells were at G1 stage. Hormone showed a promoting effect on the proliferation of granulosa cells on the 2nd day after cultivation. This promoting effect was the most obvious on the 4th day after cultivation. But no significant effects of hormone on the granulosa cell morphology and cytoskeleton were detected.

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