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Research Article |
College of Life Sciences,3 Peking University, Beijing 100871, China
Shenzhen Graduate School,4 Peking University, Shenzhen 518055, China
Beijing Laboratory Animal Research Center,5 Beijing 100083, China
Institute of Zoology,6 Chinese Academy of Science, Beijing 100871, China
ABSTRACT
Mature mouse oocytes currently can be generated in vitro from the primary oocytes of primordial follicles but not from premeiotic fetal germ cells. In this study we established a simple, efficient method that can be used to obtain mature oocytes from the premeiotic germ cells of a fetal mouse 12.5 days postcoitum (dpc). Mouse 12.5-dpc fetal ovaries were transplanted under the kidney capsule of recipient mice to initiate oocyte growth from the premeiotic germ cell stage, and they were recovered after 14 days. Subsequently, the primary and early secondary follicles generated in the ovarian grafts were isolated and cultured for 16 days in vitro. The mature oocytes ovulated from these follicles were able to fertilize in vitro to produce live offspring. We further show that the in vitro fertilization offspring were normal and able to successfully mate with both females and males, and the patterns of the methylated sites of the in vitro mature oocytes were similar to those of normal mice. This is the first report describing premeiotic fetal germ cells able to enter a second meiosis and support embryonic development to term by a combination of in vivo transplantation and in vitro culture. In addition, we have shown that the whole process of oogenesis, from premeiotic germ cells to germinal vesicle (GV)-stage oocytes, can be carried out under the kidney capsule.
developmental biology, fetal germ cells, follicular development, meiosis, mouse oogenesis, oocyte, ovary, primordial germ cell development
1 Supported by grants from the Ministry of Science and Technology (2001CB510106), Science and Technology Plan of Beijing Municipal Government (H020220050290), National Nature Science Foundation of China for Outstanding Young Scientist Award (30125022) and for Creative Research Groups (30421004), and the Bill & Melinda Gates Foundation (37871) to H.D., and the Chinese Postdoctoral Science Foundation (2005037030) to W.S.
2 Correspondence. FAX: 86 10 6275 6954; hongkui_deng{at}pku.edu.cn
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