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J Vis Exp. 2014 Aug 25;(90):e51835. doi: 10.3791/51835.

Organotypic slice cultures to study oligodendrocyte dynamics and myelination.

Author information

1
Department of Physiology and Neurobiology, University of Connecticut; Department of Neurology, Yale University School of Medicine.
2
Department of Physiology and Neurobiology, University of Connecticut.
3
Department of Physiology and Neurobiology, University of Connecticut; Stem Cell Institute, University of Connecticut; akiko.nishiyama@uconn.edu.

Abstract

NG2 expressing cells (polydendrocytes, oligodendrocyte precursor cells) are the fourth major glial cell population in the central nervous system. During embryonic and postnatal development they actively proliferate and generate myelinating oligodendrocytes. These cells have commonly been studied in primary dissociated cultures, neuron cocultures, and in fixed tissue. Using newly available transgenic mouse lines slice culture systems can be used to investigate proliferation and differentiation of oligodendrocyte lineage cells in both gray and white matter regions of the forebrain and cerebellum. Slice cultures are prepared from early postnatal mice and are kept in culture for up to 1 month. These slices can be imaged multiple times over the culture period to investigate cellular behavior and interactions. This method allows visualization of NG2 cell division and the steps leading to oligodendrocyte differentiation while enabling detailed analysis of region-dependent NG2 cell and oligodendrocyte functional heterogeneity. This is a powerful technique that can be used to investigate the intrinsic and extrinsic signals influencing these cells over time in a cellular environment that closely resembles that found in vivo.

PMID:
25177825
PMCID:
PMC4828019
DOI:
10.3791/51835
[Indexed for MEDLINE]
Free PMC Article
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