Large-scale production of growing oocytes in vitro from neonatal mouse ovaries
Large-scale production of growing oocytes in vitro from neonatal mouse ovaries
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DOI：
10.1387/ijdb.082607ah
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发表时间：
2009-01-01
影响因子：
0.7
通讯作者：
Ogura, Atsuo
中科院分区：
文献类型：
--
作者：
Honda, Arata;Hirose, Michiko;Ogura, Atsuo
尽管胎儿或新生哺乳动物卵巢中含有许多未生长的卵母细胞，但大多数退化，只有少数有助于成熟卵巢中的卵母细胞池。在这里，我们报告了一个无卵泡培养系统，允许大量的这些被捕卵母细胞进入体外生长期。新生小鼠的卵母细胞多达800个，相当于大型动物的卵母细胞超过10（4）个，继续发育，形成透明的卵母细胞，并能够与精子融合。部分卵母细胞达到正常有腔卵泡的大小并进入中期1，表明生长期完成。成功的关键是顺序提供必需的营养物质和生长因子的卵母细胞，同时防止凋亡，通常发生在大多数生长的卵母细胞在体内。重要的是，母体基因组印记，这是必要的正常胚胎发育，是正确地施加在他们的基因组自主。因此，被捕的原始卵母细胞可以在体外有效地拯救，并可以进行受精和随后的胚胎发育所需的形态学和基因组修饰。这种培养系统可能为开发用于研究、临床和动物学目的的从胎儿或新生儿卵巢有效生产卵母细胞的新技术提供重要的推动力。
Although fetal or neonatal mammalian ovaries contain many non-growing oocytes within primordial follicles, most degenerate and only a few contribute to the oocyte pool in the mature ovary. Here, we report a follicle-free culture system that allows a large number of these arrested oocytes to enter the growth phase in vitro. As many as 800 oocytes from a newborn mouse, corresponding to more than 10(4) oocytes in large animals, continued to develop, formed a zona pellucida, and were able to fuse with spermatozoa. Some oocytes reached the size of those in normal antral follicles and entered metaphase 1, indicating the completion of the growth phase. The key to success was the sequential provision of essential nutrients and growth factors to the oocytes, while preventing the apoptosis that normally occurs in the majority of growing oocytes in vivo. Importantly, maternal genomic imprinting, which is necessary for normal embryonic development, was imposed correctly on their genomes autonomously. Thus, arrested primordial oocytes can be rescued effectively in vitro and can undergo the morphological and genomic modifications necessary for fertilization and subsequent embryonic development. This culture system may provide a significant impetus to the development of new techniques for the efficient production of oocytes from fetal or neonatal ovaries, for research, clinical, and zoological purposes.