Development of the mechanism of mammalian oocyte generation and novel utilization using large scale oocyte production
哺乳动物卵母细胞生成机制的发展和大规模卵母细胞生产的新利用
基本信息
- 批准号:21200018
- 负责人:
- 金额:$ 18.55万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research on Innovative Areas (Research a proposed research project)
- 财政年份:2009
- 资助国家:日本
- 起止时间:2009 至 2011
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Although neonatal mammalian ovaries contain many nongrowing primordial oocytes, most degenerate and only a few contribute to the oocyte pool in the mature ovary. If these arrested neonatal primordial oocytes could be rescued effectively, they would form a potentially valuable source for research, clinical, and zoological purposes. In this research, we developed a follicle-free culture system that enables a large number of arrested oocytes to develop in vitro. By selective culture of ovarian cells from newborn (2-4 days old) mice, spherical colonies consisting exclusively of primordial oocytes and putative thecal stem cells were obtained. Upon treatment with a stem cell factor (c-kit ligand), these embedded oocytes were released from the colonies. The released oocytes-more than 800 per animal-continued to develop without any supporting cells, formed a zona pellucida, and were able to fuse with spermatozoa. This culture system provides a unique experimental model for studying the mechanisms of oogenesis, as it can supply many granulosa cell-free oocytes at specific stages. We successfully generated MII oocytes using ectopic transplantation method that was constructed by in vitro cultured oocytes and neonatal ovarian somatic cells. Furthermore, we noticed a novel oocyte apoptosis mechanism which is controlled by oocyte autonomously.
虽然新生哺乳动物卵巢含有许多未发育的原始卵母细胞,但大多数退化,只有少数有助于成熟卵巢中的卵母细胞池。如果这些被捕的新生原始卵母细胞可以有效地拯救,他们将形成一个潜在的有价值的来源,为研究,临床和动物学的目的。在本研究中,我们开发了一种无卵泡培养系统,使大量的停滞卵母细胞在体外发育。通过选择性培养新生小鼠(2-4天)的卵巢细胞,获得了仅由原始卵母细胞和推定的卵泡膜干细胞组成的球形集落。用干细胞因子(c-kit配体)处理后,这些包埋的卵母细胞从集落中释放出来。释放的卵母细胞(每只动物超过800个)在没有任何支持细胞的情况下继续发育,形成透明的卵母细胞,并能够与精子融合。该培养体系可在特定阶段提供大量无颗粒细胞卵母细胞,为研究卵子发生机制提供了一个独特的实验模型。利用体外培养的卵母细胞和新生儿卵巢体细胞构建的异位移植法成功获得MII期卵母细胞。此外,我们还发现了一种新的由卵母细胞自主调控的卵母细胞凋亡机制。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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HONDA Arata其他文献
HONDA Arata的其他文献
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{{ truncateString('HONDA Arata', 18)}}的其他基金
Establishment of the rabbit pluripotent stem cell systems for compensating for the difference between human and mouse stem cells.
建立兔多能干细胞系统以补偿人和小鼠干细胞之间的差异。
- 批准号:
21700453 - 财政年份:2009
- 资助金额:
$ 18.55万 - 项目类别:
Grant-in-Aid for Young Scientists (B)
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