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玻璃化冷冻、胞浆内单精子注射和胚胎移植后家猫卵母细胞的体内存活。

In vivo survival of domestic cat oocytes after vitrification, intracytoplasmic sperm injection and embryo transfer.

机构信息

Audubon Center for Research of Endangered Species, New Orleans, Louisiana, USA.

出版信息

Theriogenology. 2012 Feb;77(3):531-8. doi: 10.1016/j.theriogenology.2011.08.028. Epub 2011 Oct 19.

Abstract

We evaluated: (1) cleavage rate after IVF or intracytoplasmic sperm injection (ICSI) of in vivo- and in vitro-matured oocytes after vitrification (experiment 1); and (2) fetal development after transfer of resultant ICSI-derived embryos into recipients (experiment 2). In vivo-matured cumulus-oocyte complexes (COCs) were recovered from gonadotropin-treated donors at 24 h after LH treatment. In vitro-matured oocytes were obtained by mincing ovaries (from local veterinary clinics) and placing COCs into maturation medium for 24 h. Mature oocytes were denuded and cryopreserved in a vitrification solution of 15% DMSO, 15% ethylene glycol, and 18% sucrose. In experiment 1, for both in vivo- and in vitro-matured oocytes, cleavage frequencies after IVF of control and vitrified oocytes and after ICSI of vitrified oocytes were not different (P > 0.05). After vitrification, blastocyst development occurred only in IVF-derived, in vitro-matured oocytes. In experiment 2, 18 presumptive zygotes and four two-cell embryos derived by ICSI of vitrified in vitro-matured oocytes and 19 presumptive zygotes produced from seven in vivo- and 12 in vitro-matured oocytes were transferred by laparoscopy into the oviducts of two recipients, respectively. On Day 21, there were three fetuses in one recipient and one fetus in the other. On Days 63 and 66 of gestation, four live kittens were born. In vivo viability of zygotes and/or embryos produced via ICSI of vitrified oocytes was established by birth of live kittens after transfer to recipients.

摘要

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(1)玻璃化冷冻后的体内和体外成熟卵母细胞在体外受精(IVF)或胞浆内单精子注射(ICSI)后的卵裂率(实验 1);(2)ICSI 衍生胚胎移植到受体后的胎儿发育情况(实验 2)。在 LH 处理后 24 小时,从促性腺激素处理的供体中回收体内成熟的卵丘-卵母细胞复合物(COC)。体外成熟的卵母细胞通过切碎卵巢(来自当地兽医诊所)并将 COC 放入成熟培养基中 24 小时获得。成熟的卵母细胞被剥除并保存在 15% DMSO、15%乙二醇和 18%蔗糖的玻璃化溶液中。在实验 1 中,对于体内和体外成熟的卵母细胞,对照组和玻璃化冷冻卵母细胞的 IVF 卵裂率以及玻璃化冷冻卵母细胞的 ICSI 卵裂率没有差异(P>0.05)。玻璃化后,仅在 IVF 衍生的体外成熟卵母细胞中发生囊胚发育。在实验 2 中,通过 ICSI 将 18 个推定的受精卵和 4 个二细胞胚胎从玻璃化的体外成熟卵母细胞中衍生出来,通过 7 个体内成熟和 12 个体外成熟卵母细胞产生的 19 个推定的受精卵分别通过腹腔镜转移到两个受体的输卵管中。第 21 天,一个受体中有三个胎儿,另一个受体中有一个胎儿。在妊娠第 63 天和第 66 天,四只活小猫出生。通过将玻璃化的卵母细胞的 ICSI 产生的受精卵或胚胎转移到受体中,并诞生了活小猫,证实了体内胚胎的活力。

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