Using Acutely Dissociated and Purified Oligodendrocyte Precursor Cells for High‐Throughput Drug Screening to Identify Compounds that Promote Oligodendrocyte Differentiation. Issue 1 (19th June 2018)

• Record Type: Journal Article
• Authors: Lariosa‐Willingham, Karen; Leonoudakis, Dmitri
• Editors: Bonifacino, Juan S.; Dasso, Mary; Harford, Joe B.; Lippincott‐Schwartz, Jennifer; Yamada, Kenneth M.
• Is Part Of: Current protocols in cell biology. Volume 79:Issue 1(2018)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2018-06-19
• Subjects: differentiation -- high throughput screening assay -- multiple sclerosis -- myelin basic protein -- oligodendrocyte; Cytology -- Laboratory manuals; Cytological Techniques; Cytology; Periodicals; 571.6028
• DOI: 10.1002/cpcb.49
• Languages: English
• ISSNs: 1934-2500

Abstract

Abstract: 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.