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玻璃化冷冻兔胚胎的体内发育:玻璃化装置、受体基因型和不同步的影响。

In vivo development of vitrified rabbit embryos: effects of vitrification device, recipient genotype, and asynchrony.

机构信息

Institute of Science and Animal Technology, Laboratorio de Biotecnología de la Reproducción, Universidad Politécnica de Valencia, Valencia, Spain.

出版信息

Theriogenology. 2013 Apr 15;79(7):1124-9. doi: 10.1016/j.theriogenology.2013.02.008. Epub 2013 Mar 15.

Abstract

This study was designed to evaluate the effects of vitrification device, recipient genotype, and recipient asynchrony on implantation rate, offspring rate at birth, and fetal losses of rabbit embryos. Morphologically normal embryos (N = 787) recovered at 72 hours of gestation were kept at room temperature until transfer or vitrification. Vitrified embryos in Cryotop and ministraw devices were transferred into females induced to ovulate 60 hours (asynchrony) or 72 hours (synchrony) before transfer. In addition, recipient genotypes were analyzed (maternal and paternal genotype). The number of implanted embryos was estimated by laparoscopy as number of implantation sites at day 14 of gestation. At birth, total kits born were recorded. Fetal losses were calculated as the difference between total born at birth and implanted embryos. Our data show that a combination of Cryotop device and recipient asynchrony at -12 hours provides the most successful rate of offspring at birth, although a similar implantation rate was obtained with both devices. Thus, low fetal loss rates were observed for embryos vitrified in Cryotop independently of recipient synchrony, and embryos vitrified in straws revealed a two-fold higher rate of fetal losses. Moreover, when an asynchrony between vitrified embryo and recipients was applied, higher rates of embryos developed to term were obtained regardless of the device used. Finally, we found a highly significant association of the recipient genotype with implantation rate, offspring rate at birth, and fetal losses. In conclusion, the current study findings show that Cryotop enhances offspring rate because it is associated with a lower rate of fetal loss. This study thus provides additional evidence that recipient genotype and recipient asynchrony affect offspring rate at birth and indicates that the genotype of the recipient and the recipient asynchrony have a significant effect on implantation rate and fetal losses after vitrification.

摘要

本研究旨在评估玻璃化设备、受体基因型和受体不同步对兔胚胎着床率、出生后代率和胎儿丢失率的影响。在妊娠 72 小时时回收形态正常的胚胎(N=787),并在室温下保存,直到转移或玻璃化。在 Cryotop 和 ministraw 设备中冷冻的胚胎在转移前 60 小时(不同步)或 72 小时(同步)诱导排卵的雌性体内转移。此外,还分析了受体基因型(母本和父本基因型)。通过腹腔镜在妊娠第 14 天估计着床胚胎的数量,即着床部位的数量。出生时,记录出生的总窝仔数。胎儿丢失率通过出生时的总出生数和着床胚胎数之间的差异计算。我们的数据表明,Cryotop 设备和-12 小时的受体不同步相结合提供了出生时后代最高的成功率,尽管两种设备都获得了类似的着床率。因此,Cryotop 冷冻的胚胎无论受体同步性如何,都观察到较低的胎儿丢失率,而 straws 冷冻的胚胎则显示出两倍高的胎儿丢失率。此外,当应用冷冻胚胎和受体之间的不同步时,无论使用哪种设备,获得发育到足月的胚胎的比例都更高。最后,我们发现受体基因型与着床率、出生后代率和胎儿丢失率有高度显著的相关性。总之,本研究结果表明,Cryotop 提高了后代率,因为它与较低的胎儿丢失率相关。因此,本研究进一步证明了受体基因型和受体不同步对出生后代率的影响,并表明受体的基因型和受体的不同步对玻璃化后着床率和胎儿丢失率有显著影响。

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