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%.
出处
《东北农业大学学报》
CAS
CSCD
北大核心
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)
关键词
绵羊卵巢
获卵方法
体外成熟
比较
sheep oocyte
col ecting oocytes method
in vitro maturation
comparsion