Journal of Neuroscience Methods 2013-03-30

A reliable in vitro model for studying peripheral nerve myelination in mouse.

Mark Stettner, Kathleen Wolffram, Anne K Mausberg, Christian Wolf, Sebastian Heikaus, Angelika Derksen, Thomas Dehmel, Bernd C Kieseier

Index: J. Neurosci. Methods 214(1) , 69-79, (2013)

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Abstract

The rat dorsal root ganglia (DRG) model is a long-standing in vitro model for analysis of myelination in the peripheral nervous system. For performing systematic, high throughput analysis with transgenic animals, a simplified BL6 mouse protocol is indispensable. Here we present a stable and reliable protocol for myelinating co-cultures producing a high myelin ratio using cells from C57BL/6 mice. As an easy accessible and operable method, Sudan staining proved to be efficient in myelin detection for fixed cultures. Green fatty acid stain turned out to be highly reliable for analysis of the dynamic biological processes of myelination in vital cultures. Once myelinated we were able to induce demyelination by the addition of forskolin into the model system. In addition, we provide an optimised rat DRG protocol with significantly improved myelin ratio and a comparison of the protocols presented. Our results strengthen the value of ex vivo myelination models in neurobiology.Copyright © 2013 Elsevier B.V. All rights reserved.