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Abstract To investigate in vitro surviving and proliferating characteristics of mouse spermatogonia and its ability to endure freezing-thawing procedure by single cells or in their functional structure (seminiferous tubes or testes). Kun ming mouse testes of 7 day of age were collected to isolate the seminiferous epthelial cells (spermatogonia and Sertoli cells mainly) using one step of enzymatic digestion. After being plated D-MEM, D-MEM/F12 or KSOM (each contained 10% FBS) separately, the spermatogonial survival and proliferation in vitro was tracked systematically. The seminiferous epthelial cells were stored at -70℃ and liquid nitrogen, the seminiferous tubes and testes were stored at -70℃. The results indicated mouse spermatogonia could survive and proliferate in D-MEM, D-MEM/F12. Spermatogonia decreased gradually from 1d to 9d or 1d to 8d, increased rapidly from 9d to 13d or 8d to 12d in primary culture, while the increase slowed down after 17d or 16d. Mouse spermatogania could survive a short period but couldn’t proliferate in KSOM. No visible proliferation but differentiating characteristics was observed when mouse spermatogia cultured alone. The ideal percentage of viable cells (all above 85 percent) was got after the seminiferous epthelial cells, seminiferous tubes and testes stored at -70℃ for 1 or 2 weeks or the seminiferous epthelial cells stored in liquid nitrogen for 2 weeks, 1month and 3 months.
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Received: 27 October 2006
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