Hinrichs K, DiGiorgio L M
Department of Medicine, Tufts University School of Veterinary Medicine, North Grafton, Massachusetts 01536.
J Reprod Fertil Suppl. 1991;44:369-74.
A technique was developed in which immature horse oocytes, obtained from slaughterhouse specimens, were transferred to the pre-ovulatory follicle of a mare in vivo, with resulting oocyte maturation, ovulation, fertilization and embryo development. Oocytes were collected from all follicles greater than 3 mm, and were classified as immature, maturing, expanded or denuded. The transfers were performed in the standing, tranquilized mare. The ovary containing the pre-ovulatory follicle was grasped per rectum. A trochar and cannula were placed through the abdominal wall in the flank area, ipsilateral to the grasped ovary. For the transfer, the operator introduced a needle through the cannula to puncture the outer wall of the follicle, while adjusting the position of the ovary per rectum. The mares were inseminated the day after transfer. Twenty oocyte transfers were performed. In 1 mare, 15 immature oocytes were transferred to the pre-ovulatory follicle and 12 oocytes with expanded cumuli were recovered from the follicle 24 h later. In 3 of the remaining 19 mares, the follicle filled with blood after the transfer and ovulation did not occur. Sixteen mares ovulated after oocyte transfer. One mare was killed 3 days following ovulation; flush of the removed oviduct yielded 1 embryo, 2 recently ovulated oocytes and 3 degenerating oocytes. A uterine flush for embryo recovery was performed in each of the other 15 ovulating mares, 7-11 days after ovulation. Embryos were recovered in 7 of the 15 flushes, and embryos in excess of the number of ovulations were recovered from 4 mares (2, 3, 4 and 7 embryos).
开发了一种技术，即从屠宰场标本中获取未成熟的马卵母细胞，将其体内转移至母马的排卵前卵泡，从而实现卵母细胞成熟、排卵、受精及胚胎发育。从所有直径大于3mm的卵泡中收集卵母细胞，并将其分为未成熟、成熟、扩张或裸卵。转移操作在站立、镇静的母马身上进行。通过直肠握住含有排卵前卵泡的卵巢。将套管针和套管经腹壁插入与握住的卵巢同侧的胁腹区域。进行转移时，操作人员通过套管插入一根针以穿刺卵泡外壁，同时经直肠调整卵巢位置。转移后的第二天对母马进行授精。共进行了20次卵母细胞转移。在1匹母马中，将15个未成熟卵母细胞转移至排卵前卵泡，24小时后从该卵泡中回收了12个带有扩张卵丘的卵母细胞。在其余19匹母马中的3匹中，转移后卵泡充满血液，未发生排卵。16匹母马在卵母细胞转移后排卵。1匹母马在排卵后3天被处死；对取出的输卵管冲洗后获得1个胚胎、2个刚排卵的卵母细胞和3个退化的卵母细胞。在其他15匹排卵母马中的每一匹，在排卵后7 - 11天进行子宫冲洗以回收胚胎。15次冲洗中有7次回收了胚胎，并且从4匹母马中回收的胚胎数量超过了排卵数（分别为2、3、4和7个胚胎）。
