山羊腔前卵泡卵母细胞体外发育与体外成熟的研究

IVD AND IVM OF OOCYTES FROM PREANTRAL FOLLICLES IN GOAT

通过比较 3种培养基、5个培养系统和 4种培养方法对卵母细胞生长率的影响 ,结果发现 ,就卵母细胞生长率而言 ,以MEMHEPES培养液 +5%FCS +40 μg/mlFSH +2 0ng/mlIGF- 1+2mmol/LcAMP +2mmol/L次黄嘌呤培养系统为佳。在腔前卵泡体外培养过程中 ,12个卵泡经腔期发育 ,约占同等培养条件下卵泡总数的 2 % ( 12 / 60 3)。培养卵泡经腔期发育 ,并未改善其卵母细胞的成熟率 ( 0 / 12 )。腔前卵泡卵母细胞经过不同时间的体外培养 ,发现经 12和 14d培养结束的卵母细胞中 ,有 81% ( 4 7/ 58)生发泡明显迁移至质膜下 ;培养 14d的卵母细胞中 ,约 16 4 %( 9/ 55)产生 pbI但不排出 ,因而可在胞质中见到极体 ;培养 16d得到 1枚排出 pbI的孤雌激活卵 ;同时发现培养 16～ 18d卵母细胞退化率明显增高 (P <0 0 1)。推测山羊腔前卵泡卵母细胞体外培养的适宜时间为 16d。

WT5BZ]In present experiment,M199,MEM and Waymouth MB 752/1 medium,Five culture systems(CS) and four methods(ordinary culture,collagen layer,collagen gel clot,monolayer of granulosa cell)were used for the culture of oocytes from preantral follicles in goat It appeared that MEM HEPES medium+5% FCS+40μg/ml FSH+20ng/ml IGF 1+2mmol/L cAMP+2mmol/L hypoxanthine was the best culture system in the experiment for the growth rate of oocyte During the culture of preanrtal follicles in vitro,12 follicles developed through antrum stage,accounting for 2% of the number of follicles cultured under identical condition(12/603).The maturation rate of oocytes was not improved by developing through antrum stage of follicle in culture(0/12).When oocytes from preantral follicles were cultured for different duration,81%(47/58)of oocytes occurred migration of germinal vesicle (GV)to near plasma membrane by the end of culture of 12 and 14 days In oocytes cultured for 14 days,about 16 4%(9/55)of them produced first polar bodies(pbI)which were not released.Therefore,pbI could be visible in the cytoplasm.After 16 days in culture,one parthenogenetic egg which released pbI had been obtained.At the same time,it has been found that the degeneration rate of oocyte cultured for 16 and 18 days increased abruptly(P<0 01).It is supposed that the available duration of culture of goat oocyte from preantral follicle was 16 days.