Directing human induced pluripotent stem cells into a neurosensory lineage for auditory neuron replacement
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Date
2014
Journal Title
Journal ISSN
Volume Title
Publisher
Mary Ann Liebert
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, ßIII-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.
Description
Keywords
Stem Cell, hiPSC, Differentiation, Cochlear Implant, Auditory Neuron, Deafness
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.