Generation of eggs from mouse embryonic stem cells and induced pluripotent stem cells

被引:0
作者
Katsuhiko Hayashi
Mitinori Saitou
机构
[1] Graduate School of Medicine,Department of Anatomy and Cell Biology
[2] Kyoto University,Department of Reprogramming Science
[3] Center for iPS Cell Research and Application,undefined
[4] Kyoto University,undefined
[5] Precursory Research for Embryonic Science and Technology (PRESTO),undefined
[6] Japan Science and Technology Agency,undefined
[7] Exploratory Research for Advanced Technology (ERATO),undefined
[8] Japan Science and Technology Agency,undefined
[9] The World Premier International Research Center Initiative–Institute for Integrated Cell-Material Sciences (WPI-iCeMS),undefined
[10] Kyoto University,undefined
来源
Nature Protocols | 2013年 / 8卷
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摘要
Oogenesis is an integrated process through which an egg acquires the potential for totipotency, a fundamental condition for creating new individuals. Reconstitution of oogenesis in a culture that generates eggs with proper function from pluripotent stem cells (PSCs) is therefore one of the key goals in basic biology as well as in reproductive medicine. Here we describe a stepwise protocol for the generation of eggs from mouse PSCs, such as embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs). ESCs and iPSCs are first induced into primordial germ cell–like cells (PGCLCs) that are in turn aggregated with somatic cells of female embryonic gonads, the precursors for adult ovaries. Induction of PGCLCs followed by aggregation with the somatic cells takes up to 8 d. The aggregations are then transplanted under the ovarian bursa, in which PGCLCs grow into germinal vesicle (GV) oocytes in ∼1 month. The PGCLC-derived GV oocytes can be matured into eggs in 1 d by in vitro maturation (IVM), and they can be fertilized with spermatozoa by in vitro fertilization (IVF) to obtain healthy and fertile offspring. This method provides an initial step toward reconstitution of the entire process of oogenesis in vitro.
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页码:1513 / 1524
页数:11
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