Quantifying Spiral Ganglion Neurite and Schwann Behavior on Micropatterned Polymer Substrates.
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.
Department of Otolaryngology-Head and Neck Surgery, University of Iowa Hospitals and Clinics, Iowa City, IA, USA.
Elise L
EL
Cheng
Department of Chemical and Biochemical Engineering, University of Iowa, Iowa City, IA, USA.
Braden
B
Leigh
Department of Chemical and Biochemical Engineering, University of Iowa, Iowa City, IA, USA.
C Allan
CA
Guymon
Department of Otolaryngology-Head and Neck Surgery, University of Iowa Hospitals and Clinics, Iowa City, IA, USA. marlan-hansen@uiowa.edu.|Department of Neurosurgery, University of Iowa Hospitals and Clinics, 200 Hawkins Drive, Iowa City, IA, 52242, USA. marlan-hansen@uiowa.edu.
Marlan R
MR
Hansen
D000818:Animals; D000831:Animals, Newborn; D001672:Biocompatible Materials; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D003051:Cochlea / Q000166:cytology; D051379:Mice; D016501:Neurites / Q000378:metabolism*; D051381:Rats; D012583:Schwann Cells / Q000166:cytology*; D013136:Spiral Ganglion / Q000166:cytology*; D013499:Surface Properties
D001672:Biocompatible Materials
Dissociated culture; Explant; Photopolymerization; Primary culture; Spiral ganglion; Topographical micropattern