摘要
从屠宰场搜集的小母羊(3-6月)卵巢卵母细胞(A组)、成年母羊卵巢卵母细胞(B组)及成年母羊超排后活体所取母细胞(C组)进行了体外成熟(IVM)、体外受精(IVF)及受精后早期胚胎体外培养(IVC)的比较研究。研究发现: (1)A组平均每个卵巢获取的卵母细胞数极显著高于B组(10. 6±2. 6vs2. 5±1. 5,P【 0. 01),与C组相比差异不显著(10. 6±2. 6vs11. 0±2. 0, P】 0. 05),但获取的优良可用卵丘卵母细胞复合体(COCS)数A组极显著高于B组(4. 5±2. 5vs1. 6±1. 5,P【 0. 01),却显著低于C组(4. 5±2. 5vs8. 5±2. 5,P【 0. 05); (2)A,B,C组获取的可用卵母细胞在M199±10%EGS±20ng/mLEGF培养液中培养后的体外培养成熟率分别为63. 8%, 65%和68. 3%,差异均不显著(P】 0. 05); (3)3种来源途径的可用卵母细胞体外成熟后用mDM液获能受精处理并培养于TCM 199-卵丘颗粒细胞饲养层(cc)共培养体系中的卵裂率(37. 5%, 35. 0%和40% )无显著差异; 12枚活体来源卵母细胞生产的2-8细胞胚移植给4只同期化受体,其中1只母羊怀孕并产下2只“试管”羔羊(16. 7% )。研究表明:初情期前小母羊比成年母羊可提供更多可用于体外受精的卵母细胞,进而提高雌性动物的早期繁殖潜能;对遗传性能较好的品种或个体可通过超数排卵处理后利用合适?
In this study, oocytes collected from slaughtered prepubertal goats (Group A) and adult goats (Group B) and those collected in vivo from multi-ovulation adult goats (Group C) were in vitro matured (IVM), in vitro fertilized (IVF) and in vitro cultured (IVC) after IVF. The average number of oocytes per ovary in Group A was significantly higher than that in Group B (10.6 ± 2.6 vs 2.5 ±(1.5), P<0.01), but was not significantly different from the of Group C (10.6 ± 2.6 vs 11.0 ± 2.0, P>0.05). The average number of valuable oocytes in Group A was significantly higher than that in Group B (4.5 ± 2.5 vs 6 ± 1.5, P<0.01), but significantly lower than that in Group C (4.5 ± 2.5 vs 8.5 ± 2,5, P<0.05). Maturation rate of valuable oocytes of Groups A, B and C cultured in (M 199) + 10% EGS + 20 ng/mL EGF was 63.8%, 65% and 68.3%, respectively, and there were no significant differences among the 3 groups. After the matured oocytes were fertilized with spermatozoa capacitated in mDM and co-cultured in TCM-199 with cumulus cells, the cleavage rate was 37.5%, 35.0% and 40%, respectively, for Groups A, B and C. To produce test-tube goats and measure the reasonability of the in vitro production process of goat eggs described above, 12 in vitro produced eggs of Group C were transferred into 4 synchronized recipient goats, and one recipient became pregnant and yielded 2 lambs. In conclusion, the results of this work indicate that more valuable oocytes can be obtained from prepubertal goats than adult goats for IVF, and for some breeds or individual animals with excellent heritable traits, in vivo collection of oocytes with suitable equipment is a reasonable and effective way to prepare valuable oocytes for IVF or other embryo engineering researches.
出处
《西南农业大学学报（自然科学版）》
CSCD
北大核心
2005年第2期197-200,共4页
Journal of Southwest Agricultural University
