摘要
Bovine cumulus-oocyte complexes (COC) (n = 4230) were used in this study to assess the effects of culture method, hormonal supplementation, and cumulus cell concentration on maturation, fertilization and development of resulting embryos. Five treatments were evaluated. 1) 10 COC/50-μl drops under oil in TCM 199 supplemented with 10% heat-treated fetal calf serum, follicle-stimulating hormone (0.5 μg/ml), luteinizing hormone (5 μg/ml), and estradiol–17β (1 μg/ml); 2) as in 1 without hormones; 3) as in 1 but in 3 ml TCM-199 in petri dishes without paraffin oil; 4) as in 2 but only 1 COC/50-μl drop; and 5) as in 1 but with denuded oocytes. After 24 h maturation, the frequencies of oocytes reaching metaphase II were 98, 84, 92, 93, and 87%, respectively, for the five treatments. In the same order, percentages of normal fertilization were 73, 70, 62, 81, and 62%, and the frequencies of embryos containing two or more blastomeres at 65 h postinsemination were 69, 82, 66, 51, and 43%. The same five treatments were used in a second study in which 3,199 oocytes were fertilized, allowed to cleave in vitro to the 2- to 3-cell stage (42 h postinsemination), and transferred to oviducts of sheep (one treatment/oviduct) for 4 days. The frequencies of morulae or blastocytes obtained were 28, 18, 23, 24, and 11% for the five treatments, respectively. After nonsurgical transfer to bovine recipients (n = 8) using fresh or frozen-thawed embryos, three pregnancies past 50 days were obtained. Only one went to term with the birth of a live heifer calf.
摘要译文
在本研究中使用牛卵丘 - 卵母细胞复合物（COC）（n \x3d 4230）来评估培养方法，激素补充和卵丘细胞浓度对成熟的影响，受精和胚胎发育。评估了五种治疗方法。 1）在TCM 199中添加10％COC / 50-μl的油滴，补充10％热处理的胎牛血清，卵泡刺激素（0.5μg/ ml），促黄体激素（5μg/ ml）和雌二醇-17β（1μg/ ml）; 2）如同没有荷尔蒙的1; 3）如1中但在3ml TCM-199培养皿中不含石蜡油;4）如2中但仅1 COC / 50-μl下降; 5）如1中但裸露的卵母细胞。成熟24小时后，到达中期II的卵母细胞频率分别为98％，84％，92％，93％和87％，对于五种治疗方法。按照相同的顺序，正常受精的百分比分别为73％，70％，62％，81％和62％，在接种后65小时，含有两个或更多个卵裂球的胚胎的频率为69％，82％，66％，51％和43％。在第二项研究中使用相同的五种处理，其中3，使199个卵母细胞受精，使其在体外切割至2至3细胞期（接种后42小时），并转移至绵羊的输卵管（一次处理/输卵管）4天。所获得的桑椹胚或胚细胞的频率分别为28,18,23,24和11％。在使用新鲜或冷冻解冻的胚胎非手术转移到牛接受者（n \x3d 8）后，获得了超过50天的三次怀孕。只有一个人在活着的小母牛出生时就去了。