Khatun Momena, Bhuiyan Mohammad Musharraf Uddin, Ahmed Jalal Uddin, Haque Aminul, Rahman Mohammad Bozlur, Shamsuddin Mohammed
Department of Surgery and Obstetrics, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh.
J Vet Sci. 2011 Mar;12(1):75-82. doi: 10.4142/jvs.2011.12.1.75.
Oocytes retrieval, in vitro maturation (IVM) and fertilization (IVF) efficiency are inevitable steps towards in vitro production of embryos. In the present study, these parameters were investigated in the ovaries of prepubertal (n = 31) and pubertal (n = 61) black Bengal goats obtained from a slaughterhouse. Nuclear maturation was evaluated upon aspiration and following IVM in TCM-199 (Earle's salt with L-glutamine and sodium bicarbonate) for 27 h at 39°C under 5% CO(2) in humidified air. The oocytes retrieval and efficiency (mean ± SD) per prepubertal and pubertal goats were 5.2 ± 0.6 and 6.8 ± 0.6, and 77.3 ± 0.1% and 80.5 ± 0.6%, respectively. Anaphase I - telophase I stages differed significantly (7.3 ± 0.8 vs. 2.6 ± 0.2, p < 0.05) between the two groups of goats. After IVM, the percentages of metaphase II were significantly higher (66.3 vs. 60.3, p < 0.05) in pubertal goats than in their prepubertal counterparts. The percentages of normal in vitro fertilization (IVF) in Fert-Tyrode's albumin lactate pyruvate of pubertal goat oocytes did not differ between Percoll and swim-up sperm separation methods (36.7 ± 0.9% vs. 32.7 ± 1.3%, p > 0.05). Furthermore, sperm capacitation by heparin alone or in combination with ionomycin did not lead to a significant increase in the normal fertilization rate (34.8 ± 1.7 vs. 32.2 ± 1.5%, respectively) in the oocytes of pubertal goats. In conclusion, the ovaries of pubertal black Bengal goats obtained from the slaughterhouse could be used for in vitro embryo production. However, further optimization of the IVM and IVF techniques are necessary for satisfactory in vitro embryo production.
取卵、体外成熟（IVM）和受精（IVF）效率是胚胎体外生产的必经步骤。在本研究中，对从屠宰场获取的青春期前（n = 31）和青春期（n = 61）黑色孟加拉山羊的卵巢进行了这些参数的研究。在吸出卵母细胞时以及在含有L - 谷氨酰胺和碳酸氢钠的Earle氏盐的TCM - 199中于39°C、5%二氧化碳的湿润空气中进行27小时的体外成熟后，评估核成熟情况。青春期前和青春期山羊的取卵数及效率（平均值±标准差）分别为5.2±0.6和6.8±0.6，以及77.3±0.1%和80.5±0.6%。两组山羊的减数分裂中期I - 末期I阶段存在显著差异（7.3±0.8对2.6±0.2，p < 0.05）。体外成熟后，青春期山羊的减数分裂中期II比例显著高于青春期前山羊（66.3对60.3，p < 0.05）。青春期山羊卵母细胞在Fert - Tyrode氏白蛋白乳酸丙酮酸培养基中的正常体外受精（IVF）比例在Percoll法和上浮精子分离法之间无差异（36.7±0.9%对32.7±1.3%，p > 0.05）。此外，单独使用肝素或与离子霉素联合进行精子获能并未导致青春期山羊卵母细胞的正常受精率显著提高（分别为34.8±1.7对32.2±1.5%）。总之，从屠宰场获取的青春期黑色孟加拉山羊的卵巢可用于体外胚胎生产。然而，为了获得满意的体外胚胎生产效果，需要进一步优化体外成熟和体外受精技术。
