Bechler Marie E
MRC Centre for Regenerative Medicine, MS Society Edinburgh Centre for MS Research, The University of Edinburgh, Edinburgh, UK.
Methods Mol Biol. 2019;1936:97-110. doi: 10.1007/978-1-4939-9072-6_6.
Important advances in our understanding of oligodendrocyte precursor cell biology and differentiation have stemmed from in vitro experiments using cultures of isolated primary oligodendrocyte precursor cells. To examine the process of myelination in the final stages of oligodendrocyte development, experimental systems have previously been limited to models utilizing neurons. Recent advances in three-dimensional culture systems, however, have opened the possibility to observe myelin sheath formation with only one cell type, the oligodendrocyte precursor cell. In this chapter, such a method is described for examining oligodendrocyte myelin sheath formation with isolated oligodendrocytes in the absence of neurons. This assay is ideal for gaining mechanistic insight into oligodendrocyte-specific regulation of myelin sheath formation. Oligodendrocyte heterogeneity can be readily assessed, determining whether different oligodendrocyte sources influence myelin sheath formation. As well, the direct impact of both physical and molecular cues on oligodendrocytes can be determined in this defined system. This assay extends the capability of two-dimensional oligodendrocyte cultures, permitting post-differentiation analysis of myelinating oligodendrocytes, the number of sheaths formed by individual oligodendrocytes, as well as the lengths of myelin sheaths formed.
我们对少突胶质前体细胞生物学和分化的理解取得了重要进展，这些进展源于使用分离的原代少突胶质前体细胞培养物进行的体外实验。为了研究少突胶质细胞发育最后阶段的髓鞘形成过程，此前实验系统仅限于利用神经元的模型。然而，三维培养系统的最新进展为仅用一种细胞类型——少突胶质前体细胞观察髓鞘形成开辟了可能性。在本章中，将描述一种在没有神经元的情况下用分离的少突胶质细胞检查少突胶质细胞髓鞘形成的方法。该测定法对于深入了解少突胶质细胞特异性髓鞘形成调节机制非常理想。可以很容易地评估少突胶质细胞的异质性，确定不同的少突胶质细胞来源是否会影响髓鞘形成。此外，在这个特定系统中可以确定物理和分子信号对少突胶质细胞的直接影响。该测定法扩展了二维少突胶质细胞培养的能力，允许对髓鞘形成的少突胶质细胞进行分化后分析、单个少突胶质细胞形成的髓鞘数量以及形成的髓鞘长度。
