Vaquié Adrien, Sauvain Alizée, Jacob Claire
Department of Biology, University of Fribourg, Fribourg, Switzerland.
Methods Mol Biol. 2018;1791:157-168. doi: 10.1007/978-1-4939-7862-5_12.
Modeling myelination in vitro allows mechanistic study of developmental myelination and short-term myelin maintenance, but analyses possible to carry out using currently available models are usually limited because of high cell density and the lack of separation between neurons and myelinating cells. Furthermore, regeneration studies of myelinated systems after lesion require compartmentalization of neuronal cell bodies, axons, and myelinating cells. Here we describe a compartmentalized method using microfluidics that allows live-cell imaging at the single-cell level to follow short- and long-term dynamic interactions of neurons and myelinating cells and large-scale analyses, e.g., RNA sequencing on pure or highly enriched neurons or myelinating cells, separately.
体外模拟髓鞘形成有助于对发育性髓鞘形成和短期髓鞘维持进行机制研究，但由于细胞密度高以及神经元与髓鞘形成细胞之间缺乏分离，使用现有模型进行的分析通常受到限制。此外，损伤后有髓系统的再生研究需要将神经元细胞体、轴突和髓鞘形成细胞分隔开。在此，我们描述了一种使用微流控技术的分隔方法，该方法允许在单细胞水平进行活细胞成像，以跟踪神经元与髓鞘形成细胞的短期和长期动态相互作用，并分别对纯的或高度富集的神经元或髓鞘形成细胞进行大规模分析，例如RNA测序。
