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Establishing Primary Cultures of Trunk Neural Crest Cells

机译：建立躯干神经嵴细胞的主要文化

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摘要

Neural crest cells constitute a unique population of progenitor cells with extensive stem cell capacities able to navigate throughout various environments in the embryo and are a source of multiple cell types, including neurons, glia, melanocytes, smooth muscles, endocrine cells, cardiac cells, and also skeletal and supportive tissues in the head. Neural crest cells are not restricted to the embryo but persist as well in adult tissues where they provide a reservoir of stem cells with great therapeutic promise. Many fundamental questions in cell, developmental, and stem cell biology can be addressed using this system. During the last decades there has been an increased availability of elaborated techniques, animal models, and molecular tools to tackle neural crest cell development. However, these approaches are often very challenging and difficult to establish and they are not adapted for rapid functional investigations of mechanisms driving cell migration and differentiation. In addition, they arenot adequate for collecting pure populations of neural crest cells usable in large scale analyses and for stem cell studies. Transferring and adapting the neural crest system in tissue culture may then represent an attractive alternative, opening up numerous prospects. Here we describe a simple method for establishing primary cultures of neural crest cells derived from trunk neural tubes using the avian embryo as a source of cells. This protocol is suited for producing pure populations of neural crest cells that can be processed for cytological, cellular, and functional approaches aimed at characterizing their phenotype, behavior, and potential.
机译：神经嵴细胞构成具有广泛的干细胞能力的独特祖细胞群，能够在胚胎中的各种环境中导航，并且是多种细胞类型的来源，包括神经元，胶质叶，黑素细胞，平滑肌，内分泌细胞，心脏细胞和还有头部骨骼和支持性组织。神经嵴细胞不限于胚胎，但在成年组织中持续存在，在那里它们提供具有良好治疗承诺的干细胞储层。可以使用该系统解决细胞，发育和干细胞生物学中的许多基本问题。在过去的几十年中，阐述了阐述了技术，动物模型和分子工具的可用性增加，以解决神经嵴细胞开发。然而，这些方法通常非常具有挑战性，并且难以建立，并且它们不适用于驱动细胞迁移和分化的机制的快速功能调查。此外，它们是足以收集在大规模分析和干细胞研究中可用的神经嵴细胞的纯群。然后，在组织培养中转移和调整神经嵴系统可以代表一个有吸引力的替代方案，开放许多前景。在这里，我们描述了一种使用禽胚作为细胞来源来建立从树干神经管的神经嵴细胞的主要培养的简单方法。该方案适用于生产纯净的神经嵴细胞群，可以用于旨在表征其表型，行为和潜力的细胞学，细胞和功能方法。

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  • 来源
    《Current Protocols in Cell Biology》 |2020年第1期|共25页
  • 作者

    Jean‐Loup Duband; Nioosha Nekooie‐Marnany; Sylvie Dufour;

  • 作者单位

    Institut Mondor de Recherches Biomédicales INSERM U955 Université Paris‐Est Créteil Créteil France;

    Institut Mondor de Recherches Biomédicales INSERM U955 Université Paris‐Est Créteil Créteil France;

    Institut Mondor de Recherches Biomédicales INSERM U955 Université Paris‐Est Créteil Créteil France;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 细胞生物学;
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