期刊
NATURE MEDICINE
卷 18, 期 3, 页码 413-U176出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nm.2669
关键词
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资金
- US National Institute on Aging [NIH R37-AG012279]
- Henry and Vivian Rosenberg Philanthropic Fund
- Sea Breeze Foundation
- Vincent Memorial Hospital
- NIH [F32-AG034809]
- Grants-in-Aid for Scientific Research [22591833] Funding Source: KAKEN
Germline stem cells that produce oocytes in vitro and fertilization-competent eggs in vivo have been identified in and isolated from adult mouse ovaries. Here we describe and validate a fluorescence-activated cell sorting-based protocol that can be used with adult mouse ovaries and human ovarian cortical tissue to purify rare mitotically active cells that have a gene expression profile that is consistent with primitive germ cells. Once established in vitro, these cells can be expanded for months and can spontaneously generate 35- to 50-mu m oocytes, as determined by morphology, gene expression and haploid (1n) status. Injection of the human germline cells, engineered to stably express GFP, into human ovarian cortical biopsies leads to formation of follicles containing GFP-positive oocytes 1-2 weeks after xenotransplantation into immunodeficient female mice. Thus, ovaries of reproductive-age women, similar to adult mice, possess rare mitotically active germ cells that can be propagated in vitro as well as generate oocytes in vitro and in vivo.
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