Majerus V, De Roover R, Etienne D, Kaidi S, Massip A, Dessy F, Donnay I
Université catholique de Louvain, Unité des Sciences vétérinaires, Louvain-la-Neuve, Belgium.
Theriogenology. 1999 Nov;52(7):1169-79. doi: 10.1016/S0093-691X(99)00209-5.
The possibility of producing embryos from oocytes repeatedly collected from unstimulated calves was tested, and results obtained before and after puberty were compared in the same animals. Ovum pick-up (OPU) coupled with in vitro embryo production was used on 2 sets of 7 and 9 calves, aged 7 to 10 m.o. at the start of the experiment. The oocytes were collected twice a week during a 2-m.o. period before puberty and a 1-m.o. period after puberty. Oocytes were fertilized and co-cultured with cumulus cells in modified synthetic oviduct fluid (SOF) up to Day 7 post insemination. Some Day 7 blastocysts were vitrified and transferred to recipient heifers. An average of 3.8 to 6.8 follicles was punctured per OPU session; 1.9 to 3.1 oocytes were collected, of which more than 60% were of good quality. The number of punctured follicles and collected oocytes varied between donors. Blastocyst rates of 19 to 27% were obtained for the 2 sets. Three normal calves were born from the transfer of 20 vitrified embryos. While no significant difference was observed for the first set of calves, a significant decrease in the number of punctured follicles was observed after puberty in the second set. A direct correlation was also obtained between the number of follicles punctured before and after puberty in the same animal. In conclusion, oocytes can be collected by repeated OPU in calves 7 to 10 m.o. old without affecting their growth or the onset of puberty. An average of 5 to 11 (range 0 to 16) blastocysts per donor was produced over 2 month. However, important variations were found between donors. The correlation observed for the number of follicles punctured before and after puberty suggests that this parameter is determined before puberty.
对从未经刺激的犊牛反复采集的卵母细胞生产胚胎的可能性进行了测试,并在同一动物中比较了青春期前后获得的结果。在两组7头和9头犊牛(实验开始时年龄为7至10个月)上采用了经阴道超声引导的卵泡穿刺(OPU)结合体外胚胎生产技术。在青春期前2个月和青春期后1个月的时间段内,每周对犊牛进行两次卵母细胞采集。卵母细胞受精后,在改良的合成输卵管液(SOF)中与卵丘细胞共同培养至授精后第7天。部分第7天的囊胚进行玻璃化冷冻,并移植到受体小母牛体内。每次OPU平均穿刺3.8至6.8个卵泡;采集到1.9至3.1枚卵母细胞,其中超过60%质量良好。不同供体穿刺的卵泡数量和采集的卵母细胞数量有所不同。两组的囊胚率分别为19%至27%。移植20枚玻璃化冷冻胚胎后出生了3头正常犊牛。第一组犊牛未观察到显著差异,而第二组犊牛在青春期后穿刺的卵泡数量显著减少。在同一动物中,青春期前后穿刺的卵泡数量之间也存在直接相关性。总之,7至10月龄的犊牛可以通过反复OPU采集卵母细胞,而不影响其生长或青春期的开始。在2个月的时间里,每个供体平均产生5至11枚(范围为0至16枚)囊胚。然而,不同供体之间存在重要差异。青春期前后穿刺卵泡数量的相关性表明,该参数在青春期前就已确定。