PSCDGs of mouse multipotent adult germline stem cells can enter and progress through meiosis to form haploid male germ cells in vitro
PSCDGs of mouse multipotent adult germline stem cells can enter and progress through meiosis to form haploid male germ cells in vitro
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DOI：
10.1016/j.diff.2010.08.001
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发表时间：
2010-11-01
期刊：
影响因子：
2.9
通讯作者：
Engel, Wolfgang
中科院分区：
文献类型：
--
作者：
Nolte, Jessica;Michelmann, Hans W.;Engel, Wolfgang
精原干细胞(SSCs)通过自我更新和分化为精子，为成人一生的精子发生提供了基础。SSC来源的细胞系被称为多能成人生殖系干细胞(MaGSCs)，最近被证明是多能的，并具有与胚胎干细胞(ESCs)相同的潜力。在使用维甲酸(RA)的分化方案中，基于双重选择策略，我们已经证明ESCs能够在体外进行减数分裂并产生单倍体雄性生殖细胞。使用这种分化方案，我们现在已经成功地在体外从maGSCs中产生了单倍体的雄性生殖细胞。将来自Stra8-EGFP转基因小鼠的MAGSCs分化为稳定的精原细胞，并进行进一步培养。将减数分裂后特异性启动子pRM1-DsRed导入这些细胞，以监测维甲酸(RA)诱导的单倍体雄配子分化。我们的方案是生产多能干细胞衍生配子(PSCDGs)的另一种方法，是研究哺乳动物精子发生、生殖系基因修饰和表观遗传学重编程的替代方法。如果能在人类干细胞来源的多能细胞系中复制，它也可以作为治疗男性不育的一种方法。(C)2010年国际差异化学会。爱思唯尔有限公司出版。保留所有权利。
Spermatogonial stem cells (SSCs) provide the basis for spermatogenesis throughout adult life by undergoing self-renewal and differentiation into sperm. SSC-derived cell lines called multipotent adult germline stem cells (maGSCs) were recently shown to be pluripotent and to have the same potential as embryonic stem cells (ESCs). In a differentiation protocol using retinoic acid(RA) and based on a double selection strategy, we have shown that ESCs are able to undergo meiosis and produce haploid male germ cells in vitro. Using this differentiation protocol we have now succeeded to generate haploid male germ cells from maGSCs in vitro. maGSCs derived from a Stra8-EGFP transgenic mouse line were differentiated into stable spermatogonial stages and further cultured. These cells were transfected with a post meiotic specific promoter construct Prm1-DsRed to monitor retinoic acid(RA) induced differentiation into haploid male gametes. Our protocol is another approach for the production of pluripotent stem cell derived gametes (PSCDGs) and is an alternative for the investigation of mammalian spermatogenesis, germ line gene modification and epigenetic reprogramming. If reproducible with pluripotent cell lines derived from human SSCs, it could also be used as a therapeutic approach for the treatment of male infertility. (C) 2010 International Society of Differentiation. Published by Elsevier Ltd. All rights reserved.