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牛胚胎克隆:体外成熟卵母细胞的应用。

Embryo cloning in cattle: the use of in vitro matured oocytes.

作者信息

Barnes F, Endebrock M, Looney C, Powell R, Westhusin M, Bondioli K

机构信息

Granada Biosciences, Inc., College Station, TX 77840.

出版信息

J Reprod Fertil. 1993 Mar;97(2):317-20. doi: 10.1530/jrf.0.0970317.

Abstract

In vitro matured (IVM) bovine oocytes were examined to determine their potential viability in embryo cloning. Activation competence, as monitored by pronuclear formation, increased with oocyte age. Oocytes readily formed a pronucleus when challenged with an electrical pulse 30 h after the onset of maturation. Developmental competence of IVM oocytes tended to increase with oocyte age (P = 0.079). Selection of IVM oocytes on the basis of the presence of a polar body 24 h after the onset of maturation and the size of the follicle from which the oocyte was derived improved development of nuclear transfer embryos (polar body positive 25% versus polar body negative 10%, P < 0.05; large follicle oocytes 31% versus small follicle oocytes 14%, P < 0.05). When selected, IVM oocytes were compared with in vivo matured oocytes recovered from superovulated cows and heifers; no difference was detected for the frequency of embryos produced, pregnancies confirmed between days 50 and 60 of gestation, or the number of calves born. We conclude that selected IVM oocytes are equivalent to in vivo matured oocytes when used for bovine embryo cloning.

摘要

对体外成熟(IVM)的牛卵母细胞进行检查,以确定其在胚胎克隆中的潜在活力。通过原核形成监测的激活能力随卵母细胞年龄增加。成熟开始30小时后,卵母细胞受到电脉冲刺激时很容易形成原核。IVM卵母细胞的发育能力倾向于随卵母细胞年龄增加(P = 0.079)。根据成熟开始24小时后极体的存在以及卵母细胞所来自卵泡的大小来选择IVM卵母细胞,可改善核移植胚胎的发育(极体阳性为25%,极体阴性为10%,P < 0.05;大卵泡卵母细胞为31%,小卵泡卵母细胞为14%,P < 0.05)。在进行选择时,将IVM卵母细胞与从超排奶牛和小母牛回收的体内成熟卵母细胞进行比较;在产生的胚胎频率、妊娠第50至60天确认的妊娠情况或出生的犊牛数量方面未检测到差异。我们得出结论,当用于牛胚胎克隆时,选择的IVM卵母细胞与体内成熟卵母细胞相当。

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