Piltti K, Lindeberg H, Aalto J, Korhonen H
Institute of Applied Biotechnology, University of Kuopio, P.O. Box 1627, FIN-70211, Kuopio, Finland.
Theriogenology. 2004 Apr 1;61(5):811-20. doi: 10.1016/j.theriogenology.2003.06.001.
The Open Pulled Straw (OPS) method of vitrification has been used successfully for cryopreserving embryos of most domestic animal species. However, there is no report of a successful delivery of offspring after transfer of vitrified embryos in carnivores, even though vitrification has been a successful freezing method for species like swine whose embryos are known to be susceptible to chilling injury. Morulae and blastocysts of farmed European polecat (Mustela putorius) were vitrified and warmed before in vitro culture in modified synthetic oviductal fluid (SOF) for a period from a few hours up to 3 days before being transferred to recipients. Survival rate after vitrification, warming and in vitro culture was 51% (50/98). A total of 50 embryos were transferred surgically into the uteri of four anesthetized recipients. Two recipients delivered a total of eight offspring (2 and 6 each) for an overall survival rate of 16% (eight live cubs/50 transferred embryos). According to our knowledge, these offspring are the first carnivores produced by transfer of in vivo embryos after vitrification by OPS. Based on the present results, we suggest that OPS vitrification can be used as an alternative cryopreservation method for mustelid embryos with pup results comparable to conventional slow freezing.
开放式拉管(OPS)玻璃化方法已成功用于冷冻保存大多数家畜物种的胚胎。然而,尽管玻璃化是一种成功的冷冻方法,例如已知胚胎易受冷害的猪等物种,但关于食肉动物玻璃化胚胎移植后成功产下后代的报道却没有。养殖的欧洲艾鼬(Mustela putorius)的桑椹胚和囊胚在改良的合成输卵管液(SOF)中进行体外培养之前,先进行玻璃化和复温,培养时间从数小时到3天不等,然后再移植到受体体内。玻璃化、复温和体外培养后的存活率为51%(50/98)。总共50枚胚胎通过手术移植到4只麻醉受体的子宫中。两名受体总共产下了8只后代(分别为2只和6只),总体存活率为16%(8只活幼崽/50枚移植胚胎)。据我们所知,这些后代是通过OPS玻璃化体内胚胎移植产生的首批食肉动物。基于目前的结果,我们建议OPS玻璃化可作为艾鼬胚胎的一种替代冷冻保存方法,其幼崽产出结果与传统慢速冷冻相当。