发现成年小鼠和人的卵巢有卵原干细胞，挑战了长达半个多世纪的卵巢生物学中心法则，即：雌性哺乳动物出生后，其卵巢不能再产生新的卵泡。这一生殖生物学新发现迫使我们重新认识卵子、卵泡的发生和卵巢的功能，检讨当前治疗卵巢不育的一些方法，同时为建立家畜全能干细胞指引了一个新方向。家畜全能干细胞至今没有突破，卵原干细胞的研发成功，将提高我国的干细胞研究水平，使在细胞水平育种、定向分子育种和基因改造成为现实，开创一个家畜繁殖生物技术的新时代。本项目拟采用不同方法处理猪卵巢诱导卵原干细胞的自我复制、增生，结合卵巢组织体外培养、免疫组织化学染色等技术，分析、验证猪卵原干细胞标记基因，并通过干细胞特异抗体包被的磁珠来分离卵原干细胞，建立猪卵原干细胞系，体内、体外等手段和方法将用于对猪卵原干细胞分化潜能的分析和鉴定。

The recent publications of purification of oogonial stem cells from adult mouse and human ovaries have challenged the central dogma of female reproductive biology that female mammals are born with fixed number of non renewing pool of germ cells. Naturally, those innovative study evoked a huge wave of controversy in the filed of reproductive and stem cell biology. However, the facts of exist of oogonial stem cells in adult ovaries will force us to reconsider of the oogenesis in adult ovaries and the ovarian biology, and are supposed to create revolutionary changes in female infertility treatment. Oogonial stem cells will provide opportunity to develop oocytes not only for medical purpose, but also for animal production and genetic modification. The research of oogonial stem cells has experienced journey from basic facts to more evident facts. With the extension of the research from mouse and human to large animals, oogonial stem cells will be more lucid enough to cause emergence of technology for stem cells which have awesome number of application. Here we proposal to purify pig oogonial stem cells from adult ovaries. In this proposal, we are planning to use ovarian tissue culture as basic protocol to analyze the rare mitotically active cells. The cytotoxins treatment of ovaries in vitro and in vivo will be employed to induce proliferation of the progenitors in ovaries. The immunohistology in ovaries with different antibodies against germ cells marker will be used to identify the primitive germ cells. As described somewhere else, the immunomagnetic bead sorting will be used to isolate the primitive germ cells. Once established, totipotency the cell lines will examined in intraovarian transplantation in pig and xenografting in NOD-SCID mice.