Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (46): 8648-8652.doi: 10.3969/j.issn.1673-8225.2011.46.023

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Effects of body cavity-like temperature unsuitable for in vitro culture on rat type A spermatogonial cells

Deng Cheng-liang1, Wang Da-li1, Li San-hua2, Zhu Jing-jing1, Tang Xiu-jun1, Gong De-zhang1   

  1. 1Department of Plastic Surgery, Affiliated Hospital of Zunyi Medical College, Zunyi  563003, Guizhou Province, China
    2Central Laboratory of Zunyi Medical College, Zunyi 563003, Guizhou Province, China
  • Received:2011-05-16 Revised:2011-06-15 Online:2011-11-12 Published:2011-11-12
  • Contact: Wang Da-li, Chief physician, Professor, Master’s supervisor, Department of Plastic Surgery, Affiliated Hospital of Zunyi Medical College, Zunyi 563003, Guizhou Province, China daliwangzy@sina.com
  • About author:Deng Cheng-liang★, Studying for master’s degree, Department of Plastic Surgery, Affiliated Hospital of Zunyi Medical College, Zunyi 563003, Guizhou Province, China dengchengliang00@163.com
  • Supported by:

    Guizhou Province Foundation for Fostering Excellent Young Science and Technology Scholars, No.(2009)26*; Chongqing Major Science and Technology Program Construction and Science and Technology Scholars Fostering Plan, No. (2009)05*

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Abstract:

BACKGROUND: Temperature would influence the proliferation and apoptosis of spermatogonial cells cultured in vitro.
OBJECTIVE: To investigate the effects of temperature on cell cycle and proliferation-related gene c-kit and cyclinD3 of type A spermatogonial cells cultured in vitro.
METHODS: The isolated spermatogenic cells by two-step enzymatic digestion were co-cultured with sertoli cells in an incubator at 32 ℃ and 37 ℃ to observe the morphological changes and growth characteristics every day. On the 8th day of in vitro culture, type A spermatogenic cells were separated by gradient centrifugation and differential adhesion method.
RESULTS AND CONCLUSION: On the 1st day of in vitro culture, suspending spermatogenic cells which were round or oval and the adherent sertoli cells that were fusiform or irregular shape were observed in 32 ℃ group and 37 ℃ group. Starting from the 4th day, the number and growth state of spermatogenic cells were gradually decreased in the 37 ℃ group, at the 8th day, the number of spermatogenic cells in the 37 ℃ group was significantly lower than that in the 32 ℃ group (P < 0.05). Flow cytometry results showed that at the 8th day of in vitro culture, the proportion of type A spermatogonial cells at S phase in the 32 ℃ group was significantly higher than that in the 37 ℃ group (P < 0.05); the relative mRNA and protein expression levels of c-kit, cyclinD3 in type A spermatogonial cells were significantly higher in the 32 ℃ group than in the 37 ℃ group (P < 0.05). Results indicate that spermatogenic cells cultured in vitro would inhibit the proliferation of type A spermatogonial cells at body cavity-like temperature (37 ℃).

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