3-D gel culture and time-lapse video microscopy of the human vestibular nerve

Abstract

Conclusions: Human inner ear neurons have an innate regenerative capacity and can be cultured in vitro in a 3-D gel. The culture technique is valuable for experimental investigations of human inner ear neuron signaling and regeneration. Objectives: To establish a new in vitro model to study human inner ear nerve signaling and regeneration. Methods: Human superior vestibular ganglion (SVG) was harvested during translabyrinthine surgery for removal of vestibular schwannoma. After dissection tissue explants were embedded and cultured in a laminin-based 3-D matrix (Matrigel™). 3-D growth cone (GC) expansion was analyzed using time-lapse video microscopy (TLVM). Neural marker expression was appraised using immunocytochemistry with fluorescence and laser confocal microscopy. Results: Tissue explants from adult human SVG could be cultured in 3-D in a gel, indicating an innate potential for regeneration. Cultured GCs were found to expand dynamically in the gel. Growth cone expansion and axonal Schwann cell alignment were documented using TLVM. Neurons were identified morphologically and through immunohistochemical staining.

Keywords::

• Neural regeneration
• cochlear implants
• growth cone
• neuron signaling

Acknowledgments

We gratefully acknowledge the oto- and neurosurgical teams at Uppsala University Hospital for providing the unique human material as well as Associate Professor Nils Hailer (Uppsala University) supplying the valuable animal material. This study was supported by ALF grants from Uppsala University Hospital and Uppsala University and by the Foundation “Tysta Skolan,” the Swedish Deafness Foundation (HRF), the Swedish Research Council (A029 0401, A0290402), and kind private donations from Börje Runögård, Sweden. Our research is part of the European Community 7th Framework Programme on Research, Technological Development and Demonstration EU-FP7-NMP-2011-Small-5 “NANOCI,” project no. 281056.

Declaration of interest: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.