山羊小腔卵泡卵母细胞的体外受精试验

Experiment of in Vitro Fertilization of Goat Small Antral Folliculat Oocytes

抽吸法采集2～6mm山羊卵巢大腔卵泡卵母细胞后，以1．5mm间距纵横切割卵巢，回收（2mm小腔卵泡卵母细胞。随机测量其直径为129．86±18.84μm。将218枚卵丘细胞层致密完整的卵母细胞分别培养于含颗粒细胞并用20mMHEPES缓冲的①TCM199＋15％GS，②①＋FSH，LH（10μg／mL）＋17β-E2（1μg／ml。），③②＋ITS（5μs／mLInsulin，Transferrln，5ng／mLSodiumselenite），④TCM199＋15％NCS＋FSH，LH，17β—E2中培养26～28h，其在熟率为19．35％（6／31），26．53％（26／98），29．45％（43／146），20．74％（11／53）。体外成熟卵母细胞与体外获能的新鲜精子授精，受精率为32．94％（28／85），2细胞胚发育率为7．14％（2／28）。

After the follicles of 2 ~ 6mm (in diameter) were aspirated, the goat ovariantissue was dissected lengthwise and crosswise with the device of four razors arrangedprallelly at 1. 5mm intervals to recover<2mm small antral follicular oocytes. The diameterof small antra1 follicular oocytes was measured. The result was 129. 86± 18. 84μm. 218oocytes with the compact and dense cumulus cells and homogeneous ooplasm were culturedin 20mM HEPES buffered;①TCM199 + 15 %GS, ②①+ FSH 、LH (10μ/mL ) + 17β- E2(1μg/mL ), ③② + LTS (5μg/ml, Insulin, Transferrin, 5ng/mI, Sodium Selenite ), ④TCM 199 + 15 %NCS + FSH 、 LH + 17β - E2 for 26 ~ 28hs, and each medium had granu1osacells. The maturation rate were 19. 35 % (6/31 ), 26. 53 % (26/98), 29. 45 % (43/146 ) and20. 75 % (1l/53 )respectively- The matured ova were incubated with capacitated spermsfor insemination. The fertilization rate was 32. 94% (28/85 ) and the rate of 2 - celldevelopment was 7. 14 % (2/28).