Noisa Parinya, Lund Carina, Kanduri Kartiek, Lund Riikka, Lähdesmäki Harri, Lahesmaa Riitta, Lundin Karolina, Chokechuwattanalert Hataiwan, Otonkoski Timo, Tuuri Timo, Raivio Taneli
School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand.
J Cell Sci. 2014 May 1;127(Pt 9):2083-94. doi: 10.1242/jcs.145755. Epub 2014 Feb 25.
Neural crest cells are specified at the border between the neural plate and the epiderm. They are capable of differentiating into various somatic cell types, including craniofacial and peripheral nerve tissues. Notch signaling plays important roles during neurogenesis; however, its function during human neural crest development is poorly understood. Here, we generated self-renewing premigratory neural-crest-like cells (pNCCs) from human pluripotent stem cells (hPSCs) and investigated the roles of Notch signaling during neural crest differentiation. pNCCs expressed various neural-crest-specifier genes, including SLUG (also known as SNAI2), SOX10 and TWIST1, and were able to differentiate into most neural crest derivatives. Blocking Notch signaling during the pNCC differentiation suppressed the expression of neural-crest-specifier genes. By contrast, ectopic expression of activated Notch1 intracellular domain (NICD1) augmented the expression of neural-crest-specifier genes, and NICD1 was found to bind to their promoter regions. Notch activity was also required for the maintenance of the premigratory neural crest state, and the suppression of Notch signaling led to the generation of neural-crest-derived neurons. Taken together, we provide a protocol for the generation of pNCCs and show that Notch signaling regulates the formation, migration and differentiation of neural crest from hPSCs.
神经嵴细胞在神经板和表皮之间的边界处特化形成。它们能够分化为多种体细胞类型，包括颅面部和外周神经组织。Notch信号通路在神经发生过程中发挥重要作用；然而，其在人类神经嵴发育过程中的功能却知之甚少。在此，我们从人多能干细胞（hPSCs）中生成了自我更新的迁移前神经嵴样细胞（pNCCs），并研究了Notch信号通路在神经嵴分化过程中的作用。pNCCs表达多种神经嵴特异性基因，包括SLUG（也称为SNAI2）、SOX10和TWIST1，并且能够分化为大多数神经嵴衍生物。在pNCC分化过程中阻断Notch信号通路会抑制神经嵴特异性基因的表达。相比之下，激活的Notch1细胞内结构域（NICD1）的异位表达增强了神经嵴特异性基因的表达，并且发现NICD1与它们的启动子区域结合。维持迁移前神经嵴状态也需要Notch活性，抑制Notch信号通路会导致神经嵴衍生神经元的产生。综上所述，我们提供了一种生成pNCCs的方法，并表明Notch信号通路调节来自hPSCs的神经嵴的形成、迁移和分化。
