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ISDN2014_0379: Innervation of peripheral and central auditory tissues by human embryonic stem cell‐derived neurons in vitro
Author(s) -
Hyakumura T.,
Needham K.,
Dottori M.,
Nayagam B.A.
Publication year - 2015
Publication title -
international journal of developmental neuroscience
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.761
H-Index - 88
eISSN - 1873-474X
pISSN - 0736-5748
DOI - 10.1016/j.ijdevneu.2015.04.299
Subject(s) - otorhinolaryngology , library science , medicine , computer science , surgery
The loss of auditory neurons that occurs with profound hearing loss is irreversible in humans. Stem cell therapy for replacement of the lost auditory neurons thereby offers a potential to restore hearing in patients with profound deafness. We have previously shown that neurons derived from human embryonic stem cells (ESCs) by treatment with Noggin and small molecule Y27632 are electrically active, and capable of innervating rat sensory hair cells in vitro. This project aims to further investigate the formation of synaptic connections between hESC-derived neurons and target auditory tissues in vitro, as stem cells must be capable of synapse formation for functional improvement after deafness. Human ESC-derived neurons were co-cultured with their peripheral target tissue, sensory hair cells, from postnatal day 1–3 rats (n = 60), or with their central target tissue, cochlear nucleus slices, from postnatal day 9-12 rats (n = 12) in vitro for 2 weeks. Synapse formation was examined using immunofluorescence and confocal microscopy. Human ESC-derived neurons innervated the rows of sensory hair cells in all co-cultures. Immunoreactivity to pre-synaptic markers synapsin 1 and vesicular glutamate transporter-1 was found near the site of innervation, similarly to auditory neurons reinnervating the hair cells. Numerous synapsin 1-positive neurites and vesicular glutamate transporter-1-positive neurites of human ESC-derived neurons also innervated cochlear nucleus slices. Human ESC-derived neurons are capable of innervating both peripheral and central targets in vitro, and these newly formed synapses contain pre-synaptic markers, and the correct glutamatergic phenotype. This assay will allow investigation of the timing and further charactrisation of synaptic connections between ESC-derived neurons with their peripheral and central targets in vitro. Synapse formation is an essential component in the development of a stem cell therapy for profound hearing loss.