Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (2): 282-286.doi: 10.3969/j.issn.1673-8225.2012.02.021

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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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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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