Huang Miller, Miller Matthew L, McHenry Lauren K, Zheng Tina, Zhen Qiqi, Ilkhanizadeh Shirin, Conklin Bruce R, Bronner Marianne E, Weiss William A
Department of Neurology and the Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, CA 94158 USA.
Gladstone Institute of Cardiovascular Disease, San Francisco, CA 94158 USA.
Sci Rep. 2016 Jan 27;6:19727. doi: 10.1038/srep19727.
Neural crest cells (NCC) are stem cells that generate different lineages, including neuroendocrine, melanocytic, cartilage, and bone. The differentiation potential of NCC varies according to the level from which cells emerge along the neural tube. For example, only anterior "cranial" NCC form craniofacial bone, whereas solely posterior "trunk" NCC contribute to sympathoadrenal cells. Importantly, the isolation of human fetal NCC carries ethical and scientific challenges, as NCC induction typically occur before pregnancy is detectable. As a result, current knowledge of NCC biology derives primarily from non-human organisms. Important differences between human and non-human NCC, such as expression of HNK1 in human but not mouse NCC, suggest a need to study human NCC directly. Here, we demonstrate that current protocols to differentiate human pluripotent stem cells (PSC) to NCC are biased toward cranial NCC. Addition of retinoic acid drove trunk-related markers and HOX genes characteristic of a posterior identity. Subsequent treatment with bone morphogenetic proteins (BMPs) enhanced differentiation to sympathoadrenal cells. Our approach provides methodology for detailed studies of human NCC, and clarifies roles for retinoids and BMPs in the differentiation of human PSC to trunk NCC and to sympathoadrenal lineages.
神经嵴细胞（NCC）是一类干细胞，可分化为不同的细胞谱系，包括神经内分泌细胞、黑素细胞、软骨细胞和骨细胞。NCC的分化潜能因细胞沿神经管出现的水平而异。例如，只有前部的“颅部”NCC形成颅面骨，而后部的“躯干”NCC仅产生交感肾上腺细胞。重要的是，分离人类胎儿NCC面临伦理和科学挑战，因为NCC诱导通常在可检测到怀孕之前就已发生。因此，目前关于NCC生物学的知识主要来自非人类生物体。人类和非人类NCC之间的重要差异，如HNK1在人类而非小鼠NCC中的表达，表明有必要直接研究人类NCC。在此，我们证明，目前将人类多能干细胞（PSC）分化为NCC的方案偏向于颅部NCC。添加视黄酸可驱动与躯干相关的标志物和具有后部特征的HOX基因。随后用骨形态发生蛋白（BMP）处理可增强向交感肾上腺细胞的分化。我们的方法为详细研究人类NCC提供了方法，并阐明了视黄酸和BMP在人类PSC向躯干NCC和交感肾上腺谱系分化中的作用。
