Pavlok A, Motlík J, Kanka J, Fulka J
Czechoslovak Academy of Sciences, Institute of Physiology and Genetics of Farm Animals, Department of Genetics, Libĕchov.
Reprod Nutr Dev. 1989;29(5):611-6. doi: 10.1051/rnd:19890510.
Oocyte cumulus complexes were aspirated from 3 to 5 mm follicles of cows prestimulated with 2.000 IU PMSG 24 h before slaughter. Oocytes matured in culture were fertilized in vitro by heparinized freshly ejaculated or epididymal spermatozoa. The cultivation procedure for fertilized eggs was the same as that used for cultivation of oocytes. From 163 matured oocytes, 109 cleaved to the 2-cell stage 24 h after fertilization and after 6 days of cultivation, 18 developed to the late morula and 18 to the blastocyst stages. Eleven blastocyts and 1 late morula were transferred surgically to the uteri of 7 recipient heifers. Two heifers became pregnant: one delivered a bull-calf at term, while the other pregnancy resulted in abortion at the 3rd month. The examination of some embryos by transmission electron microscopy showed an almost normal morphology for most cells. The degenerated cells contained mostly electron-dense residual bodies of unknown origin.
在屠宰前24小时用2000国际单位孕马血清促性腺激素（PMSG）对奶牛进行预刺激，然后从3至5毫米的卵泡中吸出卵母细胞-卵丘复合体。体外成熟的卵母细胞用肝素处理的新鲜射出或附睾精子进行体外受精。受精卵的培养程序与卵母细胞培养程序相同。163个成熟卵母细胞中，109个在受精后24小时分裂至2细胞阶段，培养6天后，18个发育至桑葚胚晚期，18个发育至囊胚阶段。11个囊胚和1个桑葚胚晚期通过手术移植到7头受体小母牛的子宫内。2头小母牛怀孕：一头足月产下一头公牛犊，另一头在怀孕第3个月流产。通过透射电子显微镜对一些胚胎进行检查，结果显示大多数细胞形态几乎正常。退化细胞主要含有来源不明的电子致密残留小体。
