Marco Berland, Mario Frei, Oscar Peralta, Marcelo Ratto
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引用次数: 2
摘要
本研究的目的是确定不同的牛配子共孵育时间对受精和胚胎发育性能的影响。体外成熟的COCs与精子在TALP培养基中以1.5 × 10(6)个精子/ml的浓度共孵育3小时(t3, n = 362)、6小时(t6, n = 358)或18小时(t18, n = 350)。共孵育结束时,将第3次和第6次的COCs分别在不含精子的受精培养基中再孵育15和12 h。将t18的卵母细胞复合物与精子悬浮液共孵育18小时。假定受精卵培养9天,在第2、8和9天评估胚胎发育。每组30个囊胚染色,记录细胞核总数。受精卵发育为≥2细胞期的平均(±SEM)百分比为71.9±5.0;第3、6、18天,第2天分别为72.5±5.3和81.2±6.1,组间差异无统计学意义(P = .3)。第8天囊胚发育率(25.6±2.8);24.2±3.3;(3、6、18)组间差异无统计学意义(P = .4)。共孵育时间最短(t3)的囊胚胚核总数最多(P < 0.05)。
Time exposure period of bovine oocytes to sperm in relation to embryo development rate and quality.
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).