Neuron/Oligodendrocyte Myelination Coculture

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1791; p. 131
• Main Authors: Pang, Yi, Simpson, Kimberly, Miguel-Hidalgo, José Javier, Savich, Renate
• Format: Journal Article
• Language: English
• Published: United States 2018
• Subjects: Animals; Axons - metabolism; Axons - ultrastructure; Biomarkers; Cell Culture Techniques; Cell Lineage; Coculture Techniques; Fluorescent Antibody Technique; Immunohistochemistry; Mice; Myelin Sheath - metabolism; Myelin Sheath - ultrastructure; Neural Stem Cells; Neuroglia - cytology; Neuroglia - metabolism; Neurons - cytology; Neurons - metabolism; Oligodendroglia - cytology; Oligodendroglia - metabolism; Primary Cell Culture; Rabbits; Rats; Spinal Cord - cytology; Spinal Cord - metabolism; Embryonic; Cell culture; Axon; Myelination; Spinal cord; Oligodendrocyte
• ISSN: 1940-6029
• DOI: 10.1007/978-1-4939-7862-5_10

Myelination cell culture systems are useful tools for studying myelin biology and myelin-related disorders. Compared to a number of established protocols for dissociated pure oligodendrocyte (OL) culture, methods for myelination culture are limited. We recently developed a mixed neuron-glia coculture system that generates robust and efficient myelination. By optimizing cell culture conditions, dissociated neural progenitor cells from embryonic rat spinal cords develop into neurons and glial cells including profiles of oligodendrocyte (OL) lineage. Within 4 weeks, OL progenitor cells (OPC) proliferate, differentiate into mature OLs, and myelinate axons. The formation of compact myelin sheath is confirmed by electron microscopy. For morphological analysis by light microscopy, cells grown on glass coverslips are fixed and immunostained for various myelin-related proteins, including those embedded within the myelin sheath and those clustered at the node of Ranvier. Myelinated axons can be quantified readily by either manual counting or ImageJ software. The culture system may also be used for electron microscopic analysis by slightly modifying the cell culture procedure.