Analysis of the spatial and morphological characteristics of oligodendrocytes from images of in vitro culture

Abstract

Oligodendrocytes (OLs) are glial cells responsible for the formation of myelin sheaths in the central nervous system. The characteristic features of the oligodendrocyte lineage, ranging from proliferative and migratory oligodendrocyte progenitor cells (OPC) to myelinating mature OLs, can be observed in vitro cultures of OL lineage cells. Here, we introduce a method for analyzing the spatial distribution of OPCs, which reflects their capacity for proliferation and migration, and the morphological complexity of mature OLs, which reflects their capacity for myelin formation, from immunostaining images of in vitro OL cultures. Through the methods described, we have demonstrated the tendency for OPCs to cluster in an environment with epidermal growth factor (EGF), and the differing morphological complexity of mature OLs according to culture medium and duration of differentiation. • The proliferative and migratory characteristics of OPCs can be evaluated by analyzing their spatial distribution. • The myelin-forming capacity of mature OLs can be measured by analyzing their morphological complexity. • Image-based analyses may be a substitute for more convoluted experiments to assess OL function.