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为了解卵母细胞体外成熟与凋亡过程,进而提高卵母细胞体外成熟率.本文采自手术标本中卵巢及卵巢组织.吸取4～10mm卵泡.获得未成熟卵母细胞.用正交设计.于培养液中加入不同量组合的人绝经期促性腺激素(hMG)、卵泡液、颗粒细胞.在37℃.CO_2 5%培养箱中培养,并应用形态学观察和原位DNA片段末端标记(Tunel)检测卵母细胞的凋亡状况.结果:在本研究体外培养条件下能使未成熟卵母细胞成熟、受精,胚胎发育至早期胚泡(Blastocyst);以含hMG 0.15IU/ml卵泡液40%,颗粒细胞10~2/ml组合的培液培养卵母细胞的成熟率、受精率及印裂率最高,而凋亡率最低(P<0.05,P<0.005).结论:未成熟卵母细胞可在体外成熟、受精,并形成早期胚泡.
In order to understand the process of oocyte maturation and apoptosis in vitro, and then improve the oocyte maturation rate in vitro.This paper is taken from the ovarian and ovarian tissues of the surgical specimens, draw 4 ~ 10mm follicles, get immature oocytes. Human menopausal gonadotrophin (hMG), follicular fluid and granulosa cells were added into the culture medium in different amounts and cultured in 37 ℃ CO 2 5% incubator, and morphological observation and in situ DNA fragment end labeling Tunel) were used to detect the apoptosis of oocytes.Results: In this study, immature oocytes could be matured, fertilized and embryo developed to early stage blastocyst.The hCG containing 0.15MU / ml follicular fluid The rate of maturation, the rate of fertilization and the rate of cleavage were the highest, while the rate of apoptosis was the lowest (P <0.05, P <0.005) .Conclusion: The immature ovum B cells can mature, fertilize and form early blastocysts in vitro.