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Sci Rep. 2016 Jun 30;6:28218. doi: 10.1038/srep28218.

Human GV oocytes generated by mitotically active germ cells obtained from follicular aspirates.

Ding X1, Liu G2, Xu B1, Wu C1, Hui N3, Ni X4, Wang J1, Du M5,6, Teng X7, Wu J1,8,9.

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Renji Hospital Shanghai Jiaotong University School of Medicine, Key Laboratory for the Genetics of Developmental &Neuropsychiatric Disorders (Ministry of Education), Bio-X Institutes, Shanghai Jiao Tong University, Shanghai 200240, China.
The First People's Hospital of Chenzhou, Chenzhou 42300, Hunan, China.
Changhai Hospital of Second Military Medical University, Shanghai 200433, China.
Department of Physiology, Second Military Medical University, 800 Xiangyin Road, Shanghai 200433, China.
Laboratory for Reproductive Immunology, Obstetrics and Gynecology Hospital, Fudan University, Shanghai 200011, China.
Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases, Obstetrics and Gynecology Hospital, Fudan University, Shanghai, 200011, China.
Center of Reproductive medicine, Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai 200040, China.
Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education, Ningxia Medical University, Yinchuan 750004, China.
Shanghai Key Laboratory of Reproductive Medicine, Shanghai 200025, China.


Human female germline stem cells (FGSCs) have opened new opportunities for understanding human oogenesis, delaying menopause, treating infertility, and providing a new strategy for preserving fertility. However, the shortage of adult human ovaries tissues available impedes their future investigations and clinical applications. Here, we have established FGSC lines from scarce ovarian cortical tissues that exist in follicular aspirates (faFGSCs), which are produced and discarded in in vitro fertilization centers worldwide. The faFGSCs have characteristics of germline stem cells involved in the gene expression profile, growth characteristics, and a normal karyotype consistent with that of FGSCs obtained from ovarian cortexes surgically removed from patients (srFGSCs). Furthermore, faFGSCs have developmental potentials including spontaneous differentiation into oocytes under feeder-free conditions, communicating with granulosa cells by gap junctions and paracrine factors, entering meiosis after RA induction, as well as forming follicles after injection into human ovarian cortical tissues xenografted into adult immunodeficient female mice. Lastly, we developed a strategy guiding FGSCs differentiated into germinal vesicle (GV) stage oocytes in vitro and revealed their developmental mechanisms. Our study not only provides a new approach to obtain human FGSCs for medical treatment, but also opens several avenues to investigate human oogenesis in vitro.

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