Vitrification of rat embryos at various developmental stages
Vitrification of rat embryos at various developmental stages
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DOI：
10.1016/s0093-691x(02)01227-x
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发表时间：
2003-04-15
期刊：
影响因子：
2.8
通讯作者：
Kasai, M
中科院分区：
文献类型：
--
作者：
Han, MS;Niwa, K;Kasai, M
研究了发育阶段对冷冻保存大鼠胚胎存活率的影响。不同发育阶段的Wistar大鼠胚胎分别用EFS40或EFS20和EFS40两步玻璃化冷冻。加温后，胚胎的存活率通过它们的形态、它们在培养中发育到囊胚(或扩张的囊胚)的能力，或者它们在移植后发育到足月的能力来评估。玻璃化冷冻后回收的胚胎大部分(91-100%)在各个发育阶段形态正常。然而，1-细胞胚胎的发育能力很低，仅暴露在EFS40中0.5min，体外存活率就从76%下降到9%。2-细胞胚胎和囊胚的体外和体内存活率，两步玻璃化冷冻显著高于一步玻璃化冷冻。一步法获得了很高的体外存活率(94-100%)，即4-8-细胞胚胎和桑椹胚。一步法玻璃化冷冻的4-细胞胚胎(40%)和8-细胞胚胎(4%)的活体存活率较低，但与未玻璃化冷冻的新鲜胚胎的存活率相近。将I-Step法玻璃化冷冻的桑椹胚移植给受体，其体内存活率为61%，与新鲜胚胎的存活率(70%)无显著差异。这些结果表明，EFS40可以玻璃化冷冻2-细胞期至囊胚期的大鼠胚胎，桑椹胚期是该物种胚胎冷冻最可行的阶段。(C)2002 Elsevier Science Inc.保留所有权利。
The effect of developmental stage on the survival of cryopreserved rat embryos was examined. Wistar rat embryos at various developmental stages were vitrified by a 1-step method with EFS40, an ethylene glycol-based solution, or by a 2-step method with EFS20 and EFS40. After warming, the survival of the embryos was assessed by their morphology, their ability to develop to blastocysts (or expanded blastocysts for blastocysts) in culture, or their ability to develop to term after transfer. Most (91-100%) of the embryos recovered after vitrification were morphologically normal in all developmental stages. However, the developmental ability of 1-cell embryos was quite low; exposing them to EFS40 for just 0.5 min decreased the in vitro survival rate from 76 to 9%. The survival rates of 2-cell embryos and blastocysts, both in vitro and in vivo, were significantly higher with a 2-step vitrification process than with a 1-step vitrification process. Very high in vitro survival rates (94-100%) were obtained in 4- to 8-cell embryos and morulae in the 1-step method. Although survival rates in vivo of 4-cell (40%) and 8-cell (4%) embryos vitrified by the 1-step method were comparatively low, the values were similar to those obtained in non-vitrified fresh embryos. When morulae vitrified by the I-step method were transferred to recipients, the in vivo survival rate (61%) was high, and not significantly different from that of fresh embryos (70%). These results show that rat embryos at the 2-cell to blastocyst stages can be vitrified with EFS40, and that the morula stage is the most feasible stage for embryo cryopreservation in this species. (C) 2002 Elsevier Science Inc. All rights reserved.