Sekizar Sowmya, Williams Anna
MRC Centre for Regenerative Medicine and MS Society Edinburgh Centre, Edinburgh bioQuarter, The University of Edinburgh, Edinburgh, UK.
Methods Mol Biol. 2019;1936:169-183. doi: 10.1007/978-1-4939-9072-6_10.
In vitro culture systems have been invaluable in understanding the cell biology of oligodendrocytes; the monoculture of primary oligodendroglia has helped characterize different stages of oligodendrocyte maturation in the absence of neurons. However, oligodendrocyte monocultures do not model the interaction of oligodendrocytes with neurons where they form myelin wraps. To circumvent this problem, coculture systems were developed; oligodendrocytes and neurons are cultured together, facilitating the study of myelin wraps and the interaction between the two cell types. However, this coculture system also has limitations, as other cells are not present and it does not represent the three-dimensional multicellular structure seen in vivo. Some of these limitations are resolved by using ex vivo slice cultures to serve as a three-dimensional culture system that is more similar to in vivo and can be used to study myelination, demyelination, and remyelination, over extended periods of time. Slice cultures are economical compared to in vivo studies and live imaging using them is less challenging. The focus of this chapter is to describe how to culture brain and spinal cord slices of mice and use them to study myelination, demyelination, and remyelination.
体外培养系统对于理解少突胶质细胞的细胞生物学特性具有重要价值;原代少突胶质细胞的单培养有助于在无神经元的情况下表征少突胶质细胞成熟的不同阶段。然而,少突胶质细胞单培养无法模拟少突胶质细胞与形成髓鞘包裹的神经元之间的相互作用。为了解决这个问题,人们开发了共培养系统;将少突胶质细胞和神经元一起培养,便于研究髓鞘包裹以及这两种细胞类型之间的相互作用。然而,这种共培养系统也有局限性,因为不存在其他细胞,且它不能代表体内所见的三维多细胞结构。通过使用离体脑片培养作为一种更类似于体内的三维培养系统,可以解决其中一些局限性,该系统可用于长时间研究髓鞘形成、脱髓鞘和髓鞘再生。与体内研究相比,脑片培养经济实惠,并且使用它们进行实时成像的难度较小。本章的重点是描述如何培养小鼠的脑片和脊髓片,并利用它们来研究髓鞘形成、脱髓鞘和髓鞘再生。