Wiszniak Sophie
Centre for Cancer Biology, University of South Australia and SA Pathology, Adelaide, SA, Australia.
Methods Mol Biol. 2019;1976:83-95. doi: 10.1007/978-1-4939-9412-0_7.
In vitro culture of neural crest cells allows for the manipulation and study of neural crest cell function in a cell-autonomous manner. While several stable neural crest cell lines exist, the transformed nature of these cells may not closely reflect the in vivo properties of neural crest cells, hence making molecular and functional analyses using these cell lines difficult to interpret. Here we describe a robust method to culture primary mouse neural crest cells ex vivo for several days to weeks in culture. We further describe a method for siRNA knockdown in these cells to study gene function. This culture method can also be adapted for other molecular analyses, including addition of small-molecule inhibitors and/or growth factors to the culture media, as well as culturing neural crest cells from knockout or genetically modified mice.
神经嵴细胞的体外培养能够以细胞自主的方式对神经嵴细胞功能进行操作和研究。虽然存在几种稳定的神经嵴细胞系，但这些细胞的转化性质可能无法紧密反映神经嵴细胞的体内特性，因此使用这些细胞系进行分子和功能分析难以解释。在此，我们描述了一种可靠的方法，可将原代小鼠神经嵴细胞在体外培养数天至数周。我们还描述了一种在这些细胞中进行siRNA敲低以研究基因功能的方法。这种培养方法还可适用于其他分子分析，包括向培养基中添加小分子抑制剂和/或生长因子，以及培养来自基因敲除或基因修饰小鼠的神经嵴细胞。
