Faculty of Medicine, Department of Physiology, University of Helsinki, Helsinki 00014, Finland.
Inserm, Jean-Pierre Aubert Research Center, Development and Plasticity of the Neuroendocrine Brain, Unité 1172, 59045 Lille Cedex, France; School of Medicine, University of Lille, Lille 59000, France.
Stem Cell Reports. 2016 Aug 9;7(2):149-57. doi: 10.1016/j.stemcr.2016.06.007. Epub 2016 Jul 14.
Gonadotropin-releasing hormone (GnRH) neurons regulate human puberty and reproduction. Modeling their development and function in vitro would be of interest for both basic research and clinical translation. Here, we report a three-step protocol to differentiate human pluripotent stem cells (hPSCs) into GnRH-secreting neurons. Firstly, hPSCs were differentiated to FOXG1, EMX2, and PAX6 expressing anterior neural progenitor cells (NPCs) by dual SMAD inhibition. Secondly, NPCs were treated for 10 days with FGF8, which is a key ligand implicated in GnRH neuron ontogeny, and finally, the cells were matured with Notch inhibitor to bipolar TUJ1-positive neurons that robustly expressed GNRH1 and secreted GnRH decapeptide into the culture medium. The protocol was reproducible both in human embryonic stem cells and induced pluripotent stem cells, and thus provides a translational tool for investigating the mechanisms of human puberty and its disorders.
促性腺激素释放激素 (GnRH) 神经元调节人类的青春期和生殖功能。在体外模拟其发育和功能对于基础研究和临床转化都将具有重要意义。在此，我们报告了一种将人类多能干细胞 (hPSC) 分化为 GnRH 分泌神经元的三步法方案。首先，通过双重 SMAD 抑制将 hPSC 分化为表达 FOXG1、EMX2 和 PAX6 的前脑神经祖细胞 (NPC)。其次，用 FGF8 处理 NPC 10 天，FGF8 是一种与 GnRH 神经元发生相关的关键配体，最后，用 Notch 抑制剂将细胞成熟为具有双向 TUJ1 阳性的神经元，这些神经元强烈表达 GNRH1 并将 GnRH 十肽分泌到培养基中。该方案在人胚胎干细胞和诱导多能干细胞中都具有可重复性，因此为研究人类青春期及其障碍的机制提供了一种转化工具。
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