Cryotop facilitates high developmental ability of re-vitrified mouse embryos
Cryotop facilitates high developmental ability of re-vitrified mouse embryos
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发表时间：
2011
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通讯作者：
J. Ito;M. Kuramochi;A. Inoue;K. Yabe;Katsuyoshi Fujiwara;Osamu Nishikawa;K. Arishima;S. Sonoki;T. Masaoka;N. Kashiwazaki
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作者：
J. Ito;M. Kuramochi;A. Inoue;K. Yabe;Katsuyoshi Fujiwara;Osamu Nishikawa;K. Arishima;S. Sonoki;T. Masaoka;N. Kashiwazaki
众所周知，玻璃化冷冻不仅是保存生殖细胞，而且是保存生殖器官的有效手段。尽管有各种设备用于玻璃化哺乳动物胚胎，但据报道，在其他哺乳动物物种中，Cryotop方法比其他设备更好地开发玻璃化温热胚胎。然而，即使在小鼠中，关于胚胎的多轮玻璃化冷冻（再玻璃化）的信息也非常有限。本研究探讨了小鼠2-细胞期、4-细胞期、桑椹期和囊胚期胚胎在同一时期重复玻璃化冷冻对胚胎发育能力的影响。从输卵管收集卵丘-卵母细胞复合体，并与冷冻-解冻的附睾精子受精。体外受精后，胚胎培养至120 h。收集2-细胞期（24 h）、4-细胞期（47 h）、桑椹期（72 h）和囊胚期（96 h）的胚胎并通过Cryotop玻璃化冷冻。加温后，将胚胎培养2小时，然后重新玻璃化冷冻。玻璃化加热重复多达三次。我们的研究结果表明，再玻璃化冷冻三次不影响胚胎玻璃化冷冻在2-细胞，4-细胞，桑椹胚，或囊胚的发育能力。总之，结果表明，通过Cryotop对小鼠胚胎进行再玻璃化冷冻对胚胎的胚胎发育没有不利影响。
Vitrification is well known as an effective means of preserving not only germ cells but also reproductive organs. Although various devices are used to vitrify mammalian embryos, it has been reported in other mammalian species that the Cryotop method develops vitrified-warmed embryos better than other devices. However, information about multiple rounds of vitrification (re-vitrification) of embryos is very limited even in mice. In this study, we examined the effect of repetitively vitrified mouse embryos (2-cell stage, 4-cell stage, morula and blastocyst) at the same stage on the embryos’ developmental ability. Cumulus-oocyte complexes were collected from the oviducts and fertilized with frozen-thawed epididymal spermatozoa. After in vitro fertilization, the embryos were cultured up to 120 h. Embryos at the 2-cell stage (24 h), 4-cell stage (47 h), morular stage (72 h), and blastocyst stage (96 h) were collected and vitrified by Cryotop. After warming, embryos were cultured for 2 h and then re-vitrified. Vitrification-warming was repeated up to three times. Our results showed that re-vitrification up to three times did not affect the developmental ability of embryos vitrified at 2-cell, 4-cell, morulare, or blastocysts. Taken together, the results show that re-vitrification of mouse embryos by Cryotop did not have a detrimental effect on embryonic development of the embryos.