Deafness is a condition with a high prevalence worldwide, produced primarily by the loss of the sensory hair cells and their associated spiral ganglion neurons (SGNs). Of all the forms of deafness, auditory neuropathy is of particular concern. This condition, defined primarily by damage to the SGNs with relative preservation of the hair cells1, is responsible for a substantial proportion of patients with hearing impairment2. Although the loss of hair cells can be circumvented partially by a cochlear implant, no routine treatment is available for sensory neuron loss, as poor innervation limits the prospective performance of an implant3. Using stem cells to recover the damaged sensory circuitry is a potential therapeutic strategy. Here we present a protocol to induce differentiation from human embryonic stem cells (hESCs) using signals involved in the initial specification of the otic placode. We obtained two types of otic progenitors able to differentiate in vitro into hair-cell-like cells and auditory neurons that display expected electrophysiological properties. Moreover, when transplanted into an auditory neuropathy model, otic neuroprogenitors engraft, differentiate and significantly improve auditory-evoked response thresholds. These results should stimulate further research into the development of a cell-based therapy for deafness.
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This work was supported primarily by grants from Action on Hearing Loss (RNID) to M.N.R. Other support included Deafness Research UK (M.N.R and W.M.), the Wellcome Trust (088719, W.M.), Medical Research Council (P.W.A, H.D.M. and M.N.R) and ESTOOLS (P.W.A). S.L.J. was supported by a Wellcome Trust VIP award and the RNID. W.M. and S.L.J. are Royal Society university research fellows. Confocal images were taken at the Light Microscopy Facility of the Department of Biomedical Sciences, University of Sheffield. We are grateful for the advice of M. Mulheran and I. Russell on the tests of auditory function, provided at the earlier stages of this project, and to the assistance of P. Gokhale on the use of the InCell Analyzer. M.N.R. acknowledges the support and encouragement of his late parents, Noemí Luján-Ceballos and Juan Carlos Rivolta.
The authors declare no competing financial interests.
This file contains Supplementary Tables 1 and 2 and Supplementary Figures 1-15. (PDF 1936 kb)
This zipped file contains Supplementary Tables 3-10. Supplementary Table 3 contains the genes differentially upregulated in the FGF condition when compared to undifferentiated hESCs, using a threshold of 1.5fold-change. Supplementary Table 4 contains the genes differentially upregulated in the FGF condition when compared to cells in DFNB, using a threshold of 1.5fold-change. Supplementary Table 5 contains the genes differentially downregulated in the FGF condition when compared to undifferentiated hESCs, using a threshold of 1.5fold-change. Supplementary Table 6 contains the genes differentially downregulated in the FGF condition when compared to cells in DFNB, using a threshold of 1.5fold-change. Supplementary Table 7 the top Gene Ontology BP5 terms enriched in the genes upregulated by a 1.5 fold-change in FGF when compared to undifferentiated hESCs. Supplementary Table 8 the top Gene Ontology BP5 terms enriched in the genes upregulated by a 1.5 fold-change in FGF when compared to cells in DFNB. Supplementary Table 9 contains the top Gene Ontology BP5 terms enriched in the genes downregulated by a 1.5 fold-change in FGF when compared to undifferentiated hESCs. Supplementary Table 10 contains the top Gene Ontology BP5 terms enriched in the genes downregulated by a 1.5 fold-change in FGF when compared to cells in DFNB. (ZIP 913 kb)
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Chen, W., Jongkamonwiwat, N., Abbas, L. et al. Restoration of auditory evoked responses by human ES-cell-derived otic progenitors. Nature 490, 278–282 (2012) doi:10.1038/nature11415
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