Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (10): 1791-1794.doi: 10.3969/j.issn.1673-8225.2010.10.016

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Vitamin A effects on growth and proliferation of mouse spermatogonial stem cells during in vitro culture

Hu Jian-xin, Sun Zhao-lin, He Jian, Liang Shao-feng, Zhang Yong, Tan Zong-jian, Yuan Jun   

  1. Department of Urinary Surgery, Guizhou Provincial People’s Hospital, Guiyang   550002, Guizhou Province, China
  • Online:2010-03-05 Published:2010-03-05
  • Contact: He Jian, Associate chief physician, Master’s supervisor, Department of Urinary Surgery, Guizhou Provincial People’s Hospital, Guiyang 550002, Guizhou Province, China
  • About author:Hu Jian-xin, Master, Associate chief physician, Department of Urinary Surgery, Guizhou Provincial People’s Hospital, Guiyang 550002, Guizhou Province, China hjx918@163.com
  • Supported by:

    Tackle Key Program of Social Development in Guizhou Province, No. Qiankehe S zi [2007]1045*

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Abstract:

BACKGROUND: Vitamin A has important effects on growth of spermatogonial stem cells. At present, we have not found an inductive substance of promoting growth and differentiation of spermatogonial stem cells during in vitro culture.

OBJECTIVE: To investigate the effect of Vitamin A on growth and proliferation of mouse spermatogonial stem cells in vitro culture.

METHODS: Bilateral testis of Kunming male mice aged 5-7 days were sterilely collected. Spermatogonial stem cells were isolated and purified using adherence and noncontinuity Percoll density gradient centrifugation. Bilateral testis was obtained from Kunming male mice aged 12-15 days under sterile conditions. Sertoli cells were isolated and purified by using enzyme digestion method. Following adherence and polarization, Sertoli cells served as feeder layer. The spermatogonial stem cells were seeded on simple-layer Sertoli cells. We set two groups, In the experimental group, 1 g/L Vitamin A was added in the DMEM/F12. In the control group, Vitamin A was not added. Growth and proliferation of spermatogonial stem cells were measured using enzyme linked immunosorbent assay. The cell cycle of spermatogonial stem cells was determined by flow cytometry.

RESULTS AND CONCLUSION: At days 6, 9, 12 and 15 following coculture, absorbance value of spermatogonial stem cells in experimental group was faster than control group (P < 0.05 or 0.01). With prolongation of coculture, quantity of chromosome in S phase in spermatogonial stem cells was increased, and then decreased in the experimental group. Another division cycle began. Compared with experimental group, quantity of chromosome in S phase in spermatogonial stem cells was slowly increased in the control group (P < 0.05). During in vitro culture of mouse spermatogonial stem cells, Vitamin A can improve the proliferation and polarization of spermatogonial stem cells.

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