Stability of mouse oocytes at -80 °C: the role of the recrystallization of intracellular ice.
Stability of mouse oocytes at -80 °C: the role of the recrystallization of intracellular ice.
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DOI：
10.1530/rep-10-0438
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发表时间：
2011-04
期刊：
影响因子：
--
通讯作者：
Mazur P
中科院分区：
文献类型：
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作者：
Seki S;Mazur P
The germplasm of mutant mice is stored as frozen oocytes/embryos in many facilities world-wide. Their transport to and from such facilities should be easy and inexpensive with Dry Ice at −79 °C. The purpose of our study was to determine the stability of mouse oocytes with time at that temperature. The metaphase II oocytes were cryopreserved with a vitrification solution (EAFS10/10) developed by M. Kasai and colleagues. Two procedures were followed. In one, the samples were cooled at 187 °C/min to −196 °C, warmed to −80 °C, held at −80 °C for 1 hour to three months, and warmed to 25 °C at one of three rates. With the highest warming rate (2,950 °C/min), survival remained at 75% for the first month, but then slowly declined to 40% over the next two months. With the slowest warming (139 °C/min) survival was only ~5% even at 0 time at −80 °C. In the second procedure, the samples were cooled at 294 °C/min to −80 °C (without cooling to −196 °C) and held for up to 3 months before warming at 2,950 °C/min. Survival was ~90% after 7 days and dropped slowly to 35% after 3 months. We believe that small non-lethal quantities of intracellular ice formed during the cooling and that the intracellular crystals increased to a damaging size by recrystallization during the 3-months storage at −80 °C. From the practical point of view, this protocol yields sufficient stability to make it feasible to ship oocytes world-wide in Dry Ice.
