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不同培养条件和卵母细胞来源对黄牛卵母细胞 第一极体排出的影响
李东伟,章美玲,张运海,章孝荣
0
(安徽农业大学动物科技学院,合肥 230036; 阜阳师范学院生命科学学院,阜阳 236037; 安徽地方畜禽遗传资源保护与生物育种省级实验室,合肥 230036; 安徽省羊繁育工程技术研究中心,合肥 230036;安徽农业大学动物科技学院,合肥 230036; 安徽地方畜禽遗传资源保护与生物育种省级实验室,合肥 230036; 安徽省羊繁育工程技术研究中心,合肥 230036)
摘要:
研究了培养液组分浓度、培养方法和卵母细胞来源对黄牛卵母细胞第一极体(PB1)排出的影响。从屠宰场收集黄牛卵巢,使用抽吸法获取卵丘卵母细胞复合物(COCs)进行体外培养。结果表明,采用以下培养条件能够显著促进黄牛卵母细胞PB1排出:在培养液中添加10 μg·mL-1 FSH和LH、1.0 μg·mL-1 17-β E2、33.0 μg·mL-1丙酮酸钠;培养时间24 h,培养密度每50 μL培养液培养10~20枚COCs;卵巢储运温度25~37 ℃,储运时间<3 h,卵泡直径3~6 mm,卵丘细胞包裹2层以上。选择最佳的培养液组分浓度、培养方法和卵母细胞来源可获得最大PB1排出量,为下一步利用PB1进行保护物种、扩大优良母畜遗传资源的来源和植入前遗传学诊断等研究提供参考。
关键词:  黄牛  卵母细胞  第一极体(PB1)  体外培养
DOI:
基金项目:国家自然科学基金项目(31272442)资助。
Effects of different culture and oocyte conditions on the first polar body extrusion of cattle oocytes
LI Dong-wei,ZHANG Mei-ling,ZHANG Yun-hai,ZHANG Xiao-rong
(School of Animal Sciences and Technology, Anhui Agricultural University, Hefei 230036; School of Life Sciences, Fuyang Teachers College, Fuyang 236037; Anhui Provincial Laboratory for Local Livestock and Poultry Genetic Resource Conservation and Bio-Breeding, Hefei 230036; Engineering Research Center of Reproduction and Breeding in Sheep of Anhui Province, Hefei 230036;School of Animal Sciences and Technology, Anhui Agricultural University, Hefei 230036; Anhui Provincial Laboratory for Local Livestock and Poultry Genetic Resource Conservation and Bio-Breeding, Hefei 230036; Engineering Research Center of Reproduction and Breeding in Sheep of Anhui Province, Hefei 230036)
Abstract:
For investigating PB1 extrusion of cattle’s oocytes under different component concentrations of culture solution, culture methods in vitro and oocytes derivation, COCs suctioned from abattoir-derived ovarian follicles were cultured under different situations. Results showed that the culture conditions in vitro promoted PB1 extrusion of cattle’s oocytes significantly (P<0.05). The optimized components added in the culture medium were 10 μg·mL-1 FSH, 10 μg·mL-1 LH, 1.0 μg·mL-1 17-β E2 and 33.0 μg·mL-1 sodium pyruvate, and the optimized culture time in vitro was 24 h; culture density was 10-20 COCs per 50 μL of the culture solution; temperature of ovaries storage and transportation were at 25-37℃, and transportation time was less than 3 h; the ovarian follicle diameter was 3-6 mm; and the oocytes covered 2 layer cumulus cells would well matured. The maximum available PB1 provided research basis for protecting species, improving genetic resources of excellent females and preimplantation genetic diagnosis in the future.
Key words:  cattle  oocyte  polar body 1 (PB1)  in vitro culture

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