A Unified Methodological Framework for Vestibular Schwannoma Research

J Vis Exp. 2017 Jun 20:(124):55827. doi: 10.3791/55827.

Abstract

Vestibular schwannomas are the most common neoplasms of the cerebellopontine angle, making up 6-8% percent of all intracranial growths. Though these tumors cause sensorineural hearing loss in up to 95% of affected individuals, the molecular mechanisms underlying this hearing loss remain elusive. This article outlines the steps established in our laboratory to facilitate the collection and processing of various primary human tissue samples for downstream research applications integral to the study of vestibular schwannomas. Specifically, this work describes a unified methodological framework for the collection, processing, and culture of Schwann and schwannoma cells from surgical samples. This is integrated with parallel processing steps now considered essential for current research: the collection of tumor and nerve secretions, the preservation of RNA and the extraction of protein from collected tissues, the fixation of tissue for the preparation of sections, and the exposure of primary human cells to adeno-associated viruses for application to gene therapy. Additionally, this work highlights the translabyrinthine surgical approach to collect this tumor as a unique opportunity to obtain human sensory epithelium from the inner ear and perilymph. Tips to improve experimental quality are provided and common pitfalls highlighted.

Publication types

  • Video-Audio Media

MeSH terms

  • Ear Auricle / innervation*
  • Female
  • Hearing Loss, Sensorineural / pathology
  • Humans
  • Neuroma, Acoustic / metabolism*
  • Neuroma, Acoustic / pathology*
  • Primary Cell Culture / methods*
  • Schwann Cells / metabolism*
  • Schwann Cells / pathology*
  • Tumor Cells, Cultured