牛卵巢卵母细胞体外成熟的研究

Maturation in vitro of ovary oocytes on bovine

研究了卵巢采卵方法以及卵巢保存温度和时间、性周期阶段、激素和培养基对牛卵母细胞体外成熟的影响.结果表明:从屠宰场获取的卵巢,先切剖或抽吸卵泡再切割卵巢,可显著提高可用卵母细胞数.卵巢保存时间在2 h以内最好,最长不超过6 h.30～39 ℃的生理盐水保存卵巢,卵母细胞的成熟率(63.9 %)和卵裂率(34.4 %)均显著高于2～8 ℃(16.7 %、0 %)和20～29 ℃(54.1 %、31.7 %)保存的.采集卵母细胞时卵巢所处性周期阶段(卵泡期、黄体期)对卵母细胞体外成熟的影响不大(成熟率分别为69.2 %、64.3 %).M199是牛卵母细胞体外成熟理想的培养基,M199 + 50 IU/mL LH+ 100 IU/mL FSH + 1μg/mL 17β-E2和M199 + 50 IU/mL GnRH+1 μg/mL 17β-E2都是牛卵母细胞体外成熟理想的培养系统(成熟率分别为69.4 %、67.5 %,卵裂率分别为35.4 %、42.7 %).

The collected methods, preservation temperature and time, estrous cycle, hormone and culture medium on in vitro maturation of ovary oocytes on the bovine were studied in this paper. The ovaries collected from slaughter houses were surface cut or aspirated, and then deeply cut so that the available numbers of oocytes were remarkably increased. The optimal preservation time for ovaries should be within 2 h, and should not be more than 6 h. The oocytes with highest maturation and cleavage rates collected from ovaries preserved in saline at 30~39 ℃ showed significant differences between these collected from ovaries preserved at 2~8 ℃ (16.7 %, 0 %) and at 20~29 ℃ (54.1 %, 31.7 %), respectively (P<0.05). Oocytes collected from ovaries in different estrous cycle (follicular phase, luteal phase) had no influence on in vitro maturation of the oocytes (the maturation rate was 69.2 % and 64.3%, respectively). M199 was the ideal medium for bovine oocytes in vitro maturation. Both M199 + 50 IU/mL LH + 100 IU/mL FSH+1 μg/mL 17?-E2 and M199 + 50 IU/mL GnRH + 1 μg/mL 17?-E2 were also ideal culture system for in vitro maturation of bovine oocytes (the maturation rate was 69.4 % and 67.5 %; and the cleavage rate was 35.4 % and 42.7 %, respectively ).