Offspring derived from oocytes injected with rat sperm, frozen or freeze-dried without cryoprotection
Offspring derived from oocytes injected with rat sperm, frozen or freeze-dried without cryoprotection
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DOI：
10.1016/j.theriogenology.2007.07.017
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发表时间：
2007-10-15
期刊：
影响因子：
2.8
通讯作者：
Nakagata, N.
中科院分区：
文献类型：
--
作者：
Kaneko, T.;Kimura, S.;Nakagata, N.
精子保存是生物医学研究中保存遗传资源的一种有价值的技术。在本研究中，使用10 MM Tris-HCl和1 mM EDTA（TE缓冲液;无冷冻保护的简单溶液）冷冻或冻干大鼠精子。将结果与使用含有Equex STM和蛋黄的溶液冷冻的大鼠精子进行比较。通过将Wistar和Sprague-Dawley（SD）大鼠的精子单独注射到来自同一品系的卵母细胞中，对精子进行评价。在精子注射后存活的卵母细胞中，超过94%的卵母细胞在两种菌株的所有处理中受精。在Wistar大鼠中，分别有27%、20%、43%和30%的2-细胞胚胎发育为囊胚，35%、9%、11%和14%的2-细胞胚胎发育为来自注射新鲜、冷冻（Equex STM/蛋黄）、冷冻（TE缓冲液）和冻干精子的卵母细胞的后代。用相似来源的SD大鼠精子，分别有45%、14%、27%和7%的2-细胞胚胎发育为囊胚，22%、0%、14%和4%的2-细胞胚胎发育为后代。这些结果表明，使用TE缓冲液可以冷冻或冻干大鼠精子。我们的结论是，这种简单的保存方法，其中不需要冷冻保护，允许精子被有效地保存与维护其可复性。(C)2007爱思唯尔公司All rights reserved.
Sperm preservation is a valuable technique for maintaining genetic resources in biomedical research. In the present study, 10 MM Tris-HCl and 1 mM EDTA (TE buffer; a simple solution without cryoprotection), was used to freeze or freeze-dry rat sperm. The results were compared with rat sperm frozen using a solution containing Equex STM and egg yolk. Sperm from Wistar and Sprague-Dawley (SD) rats were evaluated by injecting them individually into oocytes derived from the same strain. Of the oocytes that survived after sperm injection, more than 94% were fertilized in all treatments of both strains. In the Wistar rat, 27, 20, 43, and 30% of 2-cell embryos developed to blastocysts, and 35, 9, 11, and 14% of 2-cell embryos developed to offspring from oocytes injected with fresh, frozen (Equex STM/egg yolk), frozen (TE buffer), and freeze-dried sperm, respectively. Using the analagous sources of sperm in the SD rat, 45, 14, 27, and 7 % of 2-cell embryos developed to blastocysts, and 22,0, 14, and 4% of 2-cell embryos developed to offspring. These results demonstrated that rat sperm could be frozen or freeze-dried using TE buffer. We concluded that this simple preservation method, in which cryoprotection was not required, allowed sperm to be preserved efficiently with maintenance of their fertilizing ability. (C) 2007 Elsevier Inc. All rights reserved.