Mattioli M, Bacci M L, Galeati G, Seren E
Istituto di Fisiologia Veterinaria Via Belmeloro 8/2 40126 Bologna Italy.
Theriogenology. 1989 Jun;31(6):1201-7. doi: 10.1016/0093-691x(89)90089-7.
Pig follicles 3 to 6 mm in diameter were everted and matured for 44 h. The oocytes were then collected and exposed to capacitated boar sperm purified by centrifugation in a two step (65 and 70%) Percoll gradient. Of 110 ova fixed 14 h after in vitro fertilization, 78% were penetrated and 47% were monospermic. Next, 681 oocytes were cultured in vitro for 44 h after in vitro fertilization and the 266 embryos which had reached the two- to four-cell stage were transferred into the oviducts of 12 synchronized recipient gilts. Four days later, 211 embryos (79%) were recovered by uterine flushing. 40.7% of these were at the blastocyst stage, and 20% were at the morula stage. In a final experiment, four out of eight gilts which had received 40 to 50 two- to four-cell embryos, were diagnosed pregnant 30 and 37 d after in vitro fertilization. One sow farrowed nine live piglets and one stillborn, two pregnancies were in progress, while one sow returned to estrus 47 d after in vitro fertilization. These results demonstrate that pig oocytes matured and fertilized in vitro can develop to the blastocyst stage and establish a normal pregnancy resulting in the birth of live piglets.
将直径为3至6毫米的猪卵泡外翻并使其成熟44小时。然后收集卵母细胞,并将其暴露于通过在两步(65%和70%)Percoll梯度中离心纯化的获能公猪精子中。在体外受精14小时后固定的110个卵子中,78%被穿透,47%为单精子受精。接下来,681个卵母细胞在体外受精后体外培养44小时,将达到二至四细胞阶段的266个胚胎转移到12头同期受体后备母猪的输卵管中。四天后,通过子宫冲洗回收了211个胚胎(79%)。其中40.7%处于囊胚阶段,20%处于桑葚胚阶段。在最后一项实验中,在体外受精30天和37天后,接受40至50个二至四细胞胚胎的8头后备母猪中有4头被诊断为怀孕。一头母猪产下9头活仔猪和1头死胎,有2头母猪仍在怀孕,而一头母猪在体外受精47天后恢复发情。这些结果表明,体外成熟和受精的猪卵母细胞可以发育到囊胚阶段,并建立正常妊娠,从而产下活仔猪。