绵羊卵巢不同获卵方法及体外成熟效果比较  

Comparsion of different collecting oocytes method and in vitro maturation on sheep

作　　者：宁方勇[1] 白秀娟[1] 

机构地区：[1]东北农业大学动物科学技术学院,哈尔滨150030

出　　处：《东北农业大学学报》2014年第8期65-71,78,共8页Journal of Northeast Agricultural University

基　　金：国家自然科学基金(30771538;31000990)

摘　　要：采用抽吸、切割和抽吸结合切割3种方法，从绵羊卵巢获取卵母细胞，分别比较3种方法获得A、B、C级卵母细胞的数量及耗时。结果表明，抽吸结合切割和切割法耗时分别为136.07和131.60 s，均显著高于抽吸法61.33 s（P〈0.05）；而获得可用卵（包括A、B级）以切割法最好为8个。对比不同激素剂量组合的3种培养液，均能获得81%以上成熟率，但成熟率差异不显著。成熟液：TCM199-HCO3＋10%FBS＋lμg·mL-1雌二醇＋2.2 g·mL-1 NaHCO3＋50μg·mL-1庆大霉素＋5μg·mL-1 FSH＋0.38 mmol·L-1丙酮酸钠＋1 IU·mL-1 LH＋10 mmol·L-1 Hepes可获得最高83.4%卵母细胞成熟率；绵羊卵巢保存在10-15℃、15-20℃和20-30℃条件下，所获卵母细胞成熟率都在82%以上，差异不显著；IA23187联合6-DMAP孤雌激活体外成熟绵羊卵母细胞，可获最高91.5%孤雌胚率。The paper collected oocytes from the sheep ovary by using 3 methods： the syrine aspiration,slicing and syrine aspiration combining with slicing,and then compared the oocytes number and time consuming to acquire A, B and C standard sheep oocytes. The results showed that the time consuming are 136.07 s and 131.60 s by syrine aspiration combining with slicing and slicing method, respectively.both of them are significantly higher than the time consuming by syrine aspiration,which time is 61.33 s （P〈0.05）; slicing is the best way to collect 8 available oocytes（including A and B sheep oocytes）.compared the 3 kinds medium with different hormone dose, all the maturation rate is higher above 81%,however, the difference is not significant, the maturation medium： TCM199-HCO3＋10%FBS＋l μg·mL-1 E2＋2.2 g·mL-1 NaHCO3 ＋50 μg·mL-1 Gentamycin＋5 μg·mL-1 FSH＋0.38 mmol·L-1 Na-Pyruvate 1 IU·mL-1 LH ＋10 mmol·L-1 Hepes can be obtained the highest oocytes maturation rate 83.4%;83.4%;the sheep oocytes conserve in 10-15℃, 15-20℃and 20-30℃, all the oocytes matura-tion rate is 82% above, the difference is not significant; IA23187 and 6-DMAP be used for parthenoge-netic activation, parthenogenetic activation embryo rate is the highest 91.5%.

关 键 词：绵羊卵巢 获卵方法 体外成熟 比较 

分 类 号：S826[农业科学—畜牧学]