Institut National de Santé et de Recherche Médicale (INSERM) U910, Université de Méditerranée Faculté de Médecine, Marseille, France.
Nat Protoc. 2011 Sep 22;6(10):1568-77. doi: 10.1038/nprot.2011.398.
A highly enriched population of neural crest cells (NCCs) from amniote embryos, such as from chicks, mice and humans, is desirable for experiments in fate determination. NCCs are also useful for testing the functional effects of molecular changes underlying numerous human diseases of neural crest derivatives and for investigating their potential for therapeutic compensation. This protocol details embryonic microdissection followed by neural tube explantation. Conditions favoring NCC expansion and the maintenance of their stem cell-like properties are described. Although neural crest-like cells can be derived from a number of sites in the mature organism, full potential is best ensured by their purification from their source tissue at the outset of migration. Going from embryo to established cell line takes 4 d; the first is the most labor-intensive day, but minimal intervention is required thereafter.
从羊膜动物胚胎（如鸡、鼠和人）中获得高度富集的神经嵴细胞（NCCs）群体，对于命运决定的实验是理想的。NCCs 也可用于测试许多神经嵴衍生物人类疾病相关的分子变化的功能影响，并研究其治疗补偿的潜力。本方案详细描述了胚胎的显微解剖，然后是神经管外植体。描述了有利于 NCC 扩增和维持其干细胞样特性的条件。尽管神经嵴样细胞可以从成熟生物体的许多部位衍生而来，但通过在迁移开始时从其源组织中纯化，才能最好地确保其充分潜能。从胚胎到建立的细胞系需要 4 天；第一天是最耗费体力的一天，但此后所需的干预最少。
