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Quantifying Spiral Ganglion Neurite and Schwann Behavior on Micropatterned Polymer Substrates
Journal article

Quantifying Spiral Ganglion Neurite and Schwann Behavior on Micropatterned Polymer Substrates

Elise L Cheng, Braden Leigh, C Allan Guymon and Marlan R Hansen
Methods in molecular biology (Clifton, N.J.), Vol.1427, pp.305-318
2016
DOI: 10.1007/978-1-4939-3615-1_18
PMCID: PMC5258114
PMID: 27259935

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Abstract

The first successful in vitro experiments on the cochlea were conducted in 1928 by Honor Fell (Fell, Arch Exp Zellforsch 7(1):69-81, 1928). Since then, techniques for culture of this tissue have been refined, and dissociated primary culture of the spiral ganglion has become a widely accepted in vitro model for studying nerve damage and regeneration in the cochlea. Additionally, patterned substrates have been developed that facilitate and direct neural outgrowth. A number of automated and semi-automated methods for quantifying this neurite outgrowth have been utilized in recent years (Zhang et al., J Neurosci Methods 160(1):149-162, 2007; Tapias et al., Neurobiol Dis 54:158-168, 2013). Here, we describe a method to study the effect of topographical cues on spiral ganglion neurite and Schwann cell alignment. We discuss our microfabrication process, characterization of pattern features, cell culture techniques for both spiral ganglion neurons and spiral ganglion Schwann cells. In addition, we describe protocols for reducing fibroblast count, immunocytochemistry, and methods for quantifying neurite and Schwann cell alignment.
Animals, Newborn Animals Surface Properties Cells, Cultured Rats Biocompatible Materials Cochlea - cytology Mice Neurites - metabolism Cell Culture Techniques - methods Spiral Ganglion - cytology Schwann Cells - cytology

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