Institute of Human Genetics, University of Göttingen, 37073 Göttingen, Germany.
Differentiation. 2010 Nov-Dec;80(4-5):184-94. doi: 10.1016/j.diff.2010.08.001. Epub 2010 Sep 1.
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 postmeiotic 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.
精原干细胞 (SSC) 通过自我更新和分化为精子，为成年期的精子发生提供基础。最近发现，源自 SSC 的多能成体生殖干细胞 (maGSCs) 具有多能性，与胚胎干细胞 (ESCs) 具有相同的潜能。在使用视黄酸 (RA) 的分化方案中，并基于双重选择策略，我们已经表明 ESCs 能够经历减数分裂并在体外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从 maGSCs 体外产生了单倍体雄性生殖细胞。源自 Stra8-EGFP 转基因小鼠系的 maGSCs 被分化为稳定的精原细胞阶段并进一步培养。这些细胞被转染了一个减数分裂后特异性启动子构建体 Prm1-DsRed，以监测视黄酸 (RA) 诱导的单倍体雄性配子分化。我们的方案是产生多能干细胞衍生配子 (PSCDG) 的另一种方法，也是研究哺乳动物精子发生、生殖系基因修饰和表观遗传重编程的替代方法。如果可以与源自人类 SSC 的多能细胞系重现，它也可以用作治疗男性不育症的治疗方法。
