Magnani Dario, Chandran Siddharthan, Wyllie David J A, Livesey Matthew R
Centre for Clinical Brain Sciences, The University of Edinburgh, Edinburgh, UK.
Euan MacDonald Centre, The University of Edinburgh, Edinburgh, UK.
Methods Mol Biol. 2019;1936:65-77. doi: 10.1007/978-1-4939-9072-6_4.
The in vitro generation of defined cellular populations from induced human pluripotent stem cells (iPSCs) provides the opportunity to work routinely with human material and, importantly, allows examination of material derived from patients with clinically and genetically diagnosed disorders. In this regard, the ability to derive oligodendrocytes in vitro represents an important resource to examine human oligodendrocyte-lineage cell biology in normal and disease contexts. Oligodendrocytes undergo characteristic physiological maturation during differentiation in vitro, and patch-clamp electrophysiology allows a detailed examination of maturation state and, potentially, pathologically related variations of ion channel expression and regulation. Here, we detail our methodology to generate oligodendrocyte precursor cells and oligodendrocytes and characterize them electrophysiologically.
从诱导多能干细胞(iPSC)体外生成特定细胞群体,为常规使用人类材料提供了机会,重要的是,还能对来自临床和基因诊断疾病患者的材料进行研究。在这方面,体外诱导生成少突胶质细胞的能力是研究正常和疾病背景下人类少突胶质细胞谱系细胞生物学的重要资源。少突胶质细胞在体外分化过程中经历典型的生理成熟过程,膜片钳电生理学可详细检测其成熟状态以及离子通道表达和调节方面可能与病理相关的变化。在此,我们详细介绍生成少突胶质前体细胞和少突胶质细胞的方法,并对其进行电生理学特征分析。