Stowers Institute for Medical Research, Kansas City, MO, USA.
Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, KS, USA.
Methods Mol Biol. 2022;2403:263-276. doi: 10.1007/978-1-0716-1847-9_18.
Neural crest cells (NCC) are a multipotent cell population that play an important role in vertebrate development. Often touted as the fourth-germ layer, NCC are induced at the border of the neural and non-neural ectoderm during the neurulation phase of embryogenesis. NCC undergo an epithelial to mesenchymal transition (EMT) that facilitates their delamination and migration throughout the embryo. After reaching their final destination, NCC then differentiate into numerous distinct cell types including neurons and glia, melanocytes, and craniofacial chondrocytes and osteoblasts. Research into the signals and mechanisms regulating each step of NCC development has been instrumental to our understanding of vertebrate development, evolution, and disease. However, studying the single and collective cellular dynamics of mammalian NCC migration has proven difficult due to the challenges accessing, and limitations visualizing, NCC within an embryo that develops in utero. The following chapter describes methods for studying the dynamics of cranial NCC migration in whole mouse embryos and in two-dimensional (2D) and 3D explant cultures of the neural plate, but these methods can be adapted for NCC at any axial level of the embryo.
神经嵴细胞（NCC）是一种多能细胞群体，在脊椎动物发育中起着重要作用。通常被称为第四胚层，NCC 在胚胎发生的神经胚形成阶段，在神经和非神经外胚层的边界处被诱导产生。NCC 经历上皮到间充质的转变（EMT），促进它们的分层和迁移整个胚胎。到达最终目的地后，NCC 然后分化为多种不同的细胞类型，包括神经元和神经胶质细胞、黑素细胞以及颅面软骨细胞和成骨细胞。对调节 NCC 发育每一步骤的信号和机制的研究，对于我们理解脊椎动物的发育、进化和疾病至关重要。然而，由于在子宫内发育的胚胎中难以获取和可视化 NCC，因此研究哺乳动物 NCC 迁移的单个和集体细胞动力学一直具有挑战性。以下章节描述了研究整个小鼠胚胎中颅神经嵴细胞迁移的动态以及神经板的二维（2D）和三维（3D）外植体培养的方法，但这些方法可以适应胚胎中任何轴向水平的 NCC。
