人类卵母细胞减数分裂进程及形态学研究  

TIMING OF MEIOTIC PROGRESSION AND MORPHOLOGICAL CHANGES IN HUMAN OOCYTES

作　　者：赵涵[1] 李媛[1] 高选[1] 陈子江[1] 

机构地区：[1]山东大学山东省立医院生殖医学中心,济南250021

出　　处：《解剖学报》2006年第4期479-483,共5页Acta Anatomica Sinica

基　　金：教育部高等学校博士学科点专项科研基金项目(20030422033);山东省优秀中青年科学家科研奖励基金项目(03BS150)

摘　　要：目的对人类未成熟卵母细胞进行体外培养,观察卵母细胞直径和卵周隙随卵母细胞成熟所发生的形态学改变,并按照其染色体变化确定人类卵母细胞减数分裂进程。方法穿刺收集多囊卵巢患者未成熟卵母细胞进行体外培养成熟,分别于体外培养4、8、12、16、20、24和28h进行卵母细胞直径及最大卵周隙测量,然后行染色体形态学观察并进行分析。结果未成熟卵母细胞体外培养10 h约有50%发生生发泡破裂(GVBD)恢复减数分裂;减数分裂Ⅰ(MⅠ)前中期由培养8 h的31.37%上升到16 h的45%,然后下降;MⅠ中期(Meta-MⅠ)在培养20 h进入高峰,占到40.63%,MⅠ后、末期(Ana/tel-MⅠ)维持时间相对较短,24 h后多数已进入减数分裂Ⅱ(MⅡ)成熟期,28 h未成熟卵母细胞体外培养的成熟率为68.85%。另外发现随着卵母细胞的体外成熟,其直径不会发生明显改变,但卵周隙(PVS)却在逐步增加。结论按照常规的体外培养方法可大致确定人类未成熟卵母细胞的体外成熟进程,且认为卵周隙的扩大可作为卵母细胞成熟分期的另一标志。Objective When cumulus-enclosed oocytes are isolated from ovarian follicles and cultured in vitro, they undergo germinal vesicle breakdown （GVBD） and extrude the first polar body, then remain in meiotic arrest. In this study, we examined the timing of progression of human oocytes through meiosis using in vitro maturation conditions, Measurements made include nuclear changes, oocyte diameter and the perivitelline space （PVS）. Methods Oocytes were collected from polycystic ovary patients by transvaginal immature follicle puncture （TV-IMFP） and matured in vitro. At 4, 8, 12, 16, 20, 24, and 28 h from the onset of culture, oocytes were put into culture droplets and analyzed under hght microscope to measure their diameter and the PVS width, then fixed and stained to evaluate the chromosomal status. Results About 50% of the oocytes underwent GVBD before 10 h culture, maximum chromatin condensation peaked at 8 h to 12 h, the majority of oocytes reached metaphase Ⅰ at 20 h, and soon anaphase and telophase Ⅰ progression occurred from 20 h to 24 h. Finally about 70% of oocytes extruded the first polar body by 28 h. With the maturation progressing, oocyte diameter did not change significantly; however, the perivitelline space width enlarged from 5.27 ＋ / - 0.88 μm to 17.18 ＋ / - 1.26μm during this time. Conclusion The results presented here delineate the timing of nuclear events in the human oocyte during maturation in vitro with this culture system. The size of PVS is related to human oocyte meiotic progression. Thus, the size of the perivitelline may be a useful indication of the maturation state of the ooeyte.

关 键 词：卵母细胞 卵周隙 减数分裂 体外培养 人 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]