Berland Marco, Frei Mario, Peralta Oscar, Ratto Marcelo
Laboratory of Animal Reproduction, Veterinary School, Faculty of Natural Resources, Catholic University of Temuco, P.O. Box 4780000, Temuco, Chile.
ISRN Vet Sci. 2011 Feb 24;2011:257627. doi: 10.5402/2011/257627. Print 2011.
The objective of the study was to determine the effect of different bovine gamete coincubation times on fertilization and embryo development performance. In vitro matured COCs were co-incubated with sperm at a concentration of 1.5 × 10(6) spermatozoa/ml in TALP medium for 3 hours (T 3, n = 362), 6 hours (T 6, n = 358), or 18 hours (T 18, n = 350). At the end of the coincubation period COCs from times 3 and 6 groups were post-incubated in a new well of fertilization medium without sperm for additional 15 and 12 h, respectively. Cumulus Oocyte Complexes from the T 18 were co-incubated with the sperm suspension for 18 hours. Presumptive zygotes were cultured for 9 days and embryo development was evaluated on days 2, 8, and 9. Thirty blastocysts from each group were stained and total number of nuclei was recorded. The mean (± SEM) percentages of zygotes to develop into ≥2 cell stage were 71.9 ± 5.0; 72.5 ± 5.3 and 81.2 ± 6.1 % for T 3, 6, and 18, respectively, on day 2 and they did not differ (P = .3) among groups. The mean percentage of blastocysts developed on day 8 (25.6 ± 2.8; 24.2 ± 3.3; 28.4 ± 4.2 % for T 3, 6, and 18, resp.) did not differ (P = .4) among groups. The total number of embryonic nuclei was greater (P < .05) for the blastocysts produced from the shortest co-incubation time (T 3).
本研究的目的是确定不同牛配子共孵育时间对受精和胚胎发育性能的影响。将体外成熟的卵丘-卵母细胞复合体(COCs)在TALP培养基中与浓度为1.5×10⁶精子/ml的精子共孵育3小时(T 3,n = 362)、6小时(T 6,n = 358)或18小时(T 18,n = 350)。在共孵育期结束时,3组和6组的COCs分别在不含精子的新受精培养基孔中再孵育15小时和12小时。T 18组的卵丘-卵母细胞复合体与精子悬液共孵育18小时。将推定的受精卵培养9天,并在第2、8和9天评估胚胎发育情况。对每组的30个囊胚进行染色并记录细胞核总数。在第2天,T 3、6和18组发育为≥2细胞阶段的受精卵的平均(±标准误)百分比分别为71.9±5.0%;72.5±5.3%和81.2±6.1%,各组间无差异(P = 0.3)。在第8天发育的囊胚的平均百分比(T 3、6和18组分别为25.6±2.8%;24.2±3.3%;28.4±4.2%)在各组间无差异(P = 0.4)。共孵育时间最短(T 3)产生的囊胚的胚胎细胞核总数更多(P < 0.05)。