Lariosa-Willingham Karen, Leonoudakis Dmitri
Teva Pharmaceuticals Biologics Discovery, Redwood City, California.
Curr Protoc Cell Biol. 2018 Jun;79(1):e49. doi: 10.1002/cpcb.49.
Multiple sclerosis (MS) is an autoimmune disease that involves an immune-mediated inflammatory response in the central nervous system and optic nerve resulting in demyelination and neural degeneration, the cause of which is unknown. The adult central nervous system has the capacity to remyelinate axons by generating new oligodendrocytes (OLs). To identify clinical candidate compounds that may promote remyelination, we have developed a high-throughput screening (HTS) assay to identify compounds that promote the differentiation of oligodendrocyte precursor cells (OPCs) into OLs. Using acutely dissociated and purified rat OPCs coupled with immunofluorescent image quantification, we have developed an OL differentiation assay. Building on OPC culturing techniques developed over the past 30 years, we have scaled up the isolation and purification process to generate sufficient quantities for HTS. We then describe the use of these acutely derived OPCs in an assay designed to identify compounds that promote differentiation into OLs. We have validated this assay with a known promoter of differentiation, thyroid hormone, and subsequently used the assay to screen the NIH clinical collection library (Lariosa-Willingham, et al., 2016). © 2018 by John Wiley & Sons, Inc.
多发性硬化症（MS）是一种自身免疫性疾病，涉及中枢神经系统和视神经中的免疫介导炎症反应，导致脱髓鞘和神经变性，其病因尚不清楚。成体中枢神经系统具有通过生成新的少突胶质细胞（OLs）来重新髓鞘化轴突的能力。为了鉴定可能促进髓鞘再生的临床候选化合物，我们开发了一种高通量筛选（HTS）试验，以鉴定促进少突胶质细胞前体细胞（OPCs）分化为OLs的化合物。利用急性解离和纯化的大鼠OPCs结合免疫荧光图像定量分析，我们开发了一种OL分化试验。基于过去30年发展起来的OPC培养技术，我们扩大了分离和纯化过程，以产生足够数量的细胞用于HTS。然后，我们描述了这些急性来源的OPCs在一种旨在鉴定促进分化为OLs的化合物的试验中的应用。我们已经用一种已知的分化促进剂甲状腺激素验证了该试验，随后用该试验筛选了美国国立卫生研究院临床收藏库（拉里奥萨 - 威林厄姆等人，2016年）。©2018约翰·威利父子公司。
