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Title: Directing human induced pluripotent stem cells into a neurosensory lineage for auditory neuron replacement
Authors: Gunewardene, Niliksha
Van Bergen, Nicole
Crombie, Duncan
Needham, Karina
Dottori, Mirella
Nayagam, Bryony
Keywords: Stem Cell
Cochlear Implant
Auditory Neuron
Issue Date: 2014
Publisher: Mary Ann Liebert
Citation: Gunewardene, N., Van Bergen, N., Crombie, D., Needham, K., Dottori, M. & Nayagam, B. A. (2014). Directing human induced pluripotent stem cells into a neurosensory lineage for auditory neuron replacement. BioResearch Open Access.
Abstract: Emerging therapies for sensorineural hearing loss include replacing damaged auditory neurons (AN) using stem cells. Ultimately, it is important that these replacement cells can be patient-matched to avoid immunorejection. As human induced pluripotent stem cells (hiPSCs) can be obtained directly from the patient, they offer an opportunity to generate patient-matched neurons for transplantation. Here, we used an established neural induction protocol to differentiate two hiPSC lines (iPS1 and iPS2) and one human embryonic stem cell line (hESC, H9) towards a neurosensory lineage in vitro. Immunocytochemistry and qRT-PCR were used to analyze the expression of key markers involved in AN development, at defined time points of differentiation. The hiPSC and hESC-derived neurosensory progenitors expressed the dorsal hindbrain marker (PAX7), otic placodal marker (PAX2), pro-neurosensory marker (SOX2), ganglion neuronal markers (NEUROD1, BRN3A, ISLET1, ├čIII-tubulin, Neurofilament kDa 160) and sensory AN markers (GATA3 and VGLUT1) over the time course examined. The hiPSC-and hESC-derived neurosensory progenitors had the highest expression levels of the sensory neural markers at 35 days in vitro. Furthermore, the neurons generated from this assay were found to be electrically active. Whilst all cell lines analyzed produced functional neurosensory-like progenitors, variabilities in the levels of marker expression were observed between hiPSC lines and within samples of the same cell line, when compared to the hESC controls. Overall, these findings indicate that this neural assay was capable of differentiating hiPSCs towards a neurosensory lineage, but emphasize the need for improving the consistency in the differentiation of hiPSCs into the required lineages.
URI: http://repository.bionicsinstitute.org:8080/handle/123456789/85
Appears in Collections:Bionic Hearing Research Publications

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