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


OBJECTIVE
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.
Department of Surgical Sciences, Head and Neck Surgery, Section of Otolaryngology, Uppsala University Hospital , Uppsala , Sweden.
Fredrik
F
Edin
Wei
W
Liu
Hao
H
Li
Francesca
F
Atturo
Peetra U
PU
Magnusson
Helge
H
Rask-Andersen
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007150:Immunohistochemistry; D018613:Microscopy, Confocal; D018715:Microscopy, Video / Q000379:methods*; D014725:Vestibular Nerve / Q000166:cytology*
Neural regeneration; cochlear implants; growth cone; neuron signaling