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Generation of Otic Lineages from Integration-Free Human-Induced Pluripotent Stem Cells Reprogrammed by mRNAs

Damage to the sensory hair cells and the spiral ganglion neurons of the cochlea leads to deafness. Induced pluripotent stem cells (iPSCs) are a promising tool to regenerate the cells in the inner ear that have been affected by pathology or have been lost. To facilitate the clinical application of iP...

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Autores principales: Boddy, Sarah L., Romero-Guevara, Ricardo, Ji, Ae-Ri, Unger, Christian, Corns, Laura, Marcotti, Walter, Rivolta, Marcelo N.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7068143/
https://www.ncbi.nlm.nih.gov/pubmed/32190057
http://dx.doi.org/10.1155/2020/3692937
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author Boddy, Sarah L.
Romero-Guevara, Ricardo
Ji, Ae-Ri
Unger, Christian
Corns, Laura
Marcotti, Walter
Rivolta, Marcelo N.
author_facet Boddy, Sarah L.
Romero-Guevara, Ricardo
Ji, Ae-Ri
Unger, Christian
Corns, Laura
Marcotti, Walter
Rivolta, Marcelo N.
author_sort Boddy, Sarah L.
collection PubMed
description Damage to the sensory hair cells and the spiral ganglion neurons of the cochlea leads to deafness. Induced pluripotent stem cells (iPSCs) are a promising tool to regenerate the cells in the inner ear that have been affected by pathology or have been lost. To facilitate the clinical application of iPSCs, the reprogramming process should minimize the risk of introducing undesired genetic alterations while conferring the cells the capacity to differentiate into the desired cell type. Currently, reprogramming induced by synthetic mRNAs is considered to be one of the safest ways of inducing pluripotency, as the transgenes are transiently delivered into the cells without integrating into the genome. In this study, we explore the ability of integration-free human-induced pluripotent cell lines that were reprogrammed by mRNAs, to differentiate into otic progenitors and, subsequently, into hair cell and neuronal lineages. hiPSC lines were induced to differentiate by culturing them in the presence of fibroblast growth factors 3 and 10 (FGF3 and FGF10). Progenitors were identified by quantitative microscopy, based on the coexpression of otic markers PAX8, PAX2, FOXG1, and SOX2. Otic epithelial progenitors (OEPs) and otic neuroprogenitors (ONPs) were purified and allowed to differentiate further into hair cell-like cells and neurons. Lineages were characterised by immunocytochemistry and electrophysiology. Neuronal cells showed inward Na(+) (I(Na)) currents and outward (I(k)) and inward K(+) (I(K1)) currents while hair cell-like cells had inward I(K1) and outward delayed rectifier K(+) currents, characteristic of developing hair cells. We conclude that human-induced pluripotent cell lines that have been reprogrammed using nonintegrating mRNAs are capable to differentiate into otic cell types.
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spelling pubmed-70681432020-03-18 Generation of Otic Lineages from Integration-Free Human-Induced Pluripotent Stem Cells Reprogrammed by mRNAs Boddy, Sarah L. Romero-Guevara, Ricardo Ji, Ae-Ri Unger, Christian Corns, Laura Marcotti, Walter Rivolta, Marcelo N. Stem Cells Int Research Article Damage to the sensory hair cells and the spiral ganglion neurons of the cochlea leads to deafness. Induced pluripotent stem cells (iPSCs) are a promising tool to regenerate the cells in the inner ear that have been affected by pathology or have been lost. To facilitate the clinical application of iPSCs, the reprogramming process should minimize the risk of introducing undesired genetic alterations while conferring the cells the capacity to differentiate into the desired cell type. Currently, reprogramming induced by synthetic mRNAs is considered to be one of the safest ways of inducing pluripotency, as the transgenes are transiently delivered into the cells without integrating into the genome. In this study, we explore the ability of integration-free human-induced pluripotent cell lines that were reprogrammed by mRNAs, to differentiate into otic progenitors and, subsequently, into hair cell and neuronal lineages. hiPSC lines were induced to differentiate by culturing them in the presence of fibroblast growth factors 3 and 10 (FGF3 and FGF10). Progenitors were identified by quantitative microscopy, based on the coexpression of otic markers PAX8, PAX2, FOXG1, and SOX2. Otic epithelial progenitors (OEPs) and otic neuroprogenitors (ONPs) were purified and allowed to differentiate further into hair cell-like cells and neurons. Lineages were characterised by immunocytochemistry and electrophysiology. Neuronal cells showed inward Na(+) (I(Na)) currents and outward (I(k)) and inward K(+) (I(K1)) currents while hair cell-like cells had inward I(K1) and outward delayed rectifier K(+) currents, characteristic of developing hair cells. We conclude that human-induced pluripotent cell lines that have been reprogrammed using nonintegrating mRNAs are capable to differentiate into otic cell types. Hindawi 2020-03-01 /pmc/articles/PMC7068143/ /pubmed/32190057 http://dx.doi.org/10.1155/2020/3692937 Text en Copyright © 2020 Sarah L. Boddy et al. http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Boddy, Sarah L.
Romero-Guevara, Ricardo
Ji, Ae-Ri
Unger, Christian
Corns, Laura
Marcotti, Walter
Rivolta, Marcelo N.
Generation of Otic Lineages from Integration-Free Human-Induced Pluripotent Stem Cells Reprogrammed by mRNAs
title Generation of Otic Lineages from Integration-Free Human-Induced Pluripotent Stem Cells Reprogrammed by mRNAs
title_full Generation of Otic Lineages from Integration-Free Human-Induced Pluripotent Stem Cells Reprogrammed by mRNAs
title_fullStr Generation of Otic Lineages from Integration-Free Human-Induced Pluripotent Stem Cells Reprogrammed by mRNAs
title_full_unstemmed Generation of Otic Lineages from Integration-Free Human-Induced Pluripotent Stem Cells Reprogrammed by mRNAs
title_short Generation of Otic Lineages from Integration-Free Human-Induced Pluripotent Stem Cells Reprogrammed by mRNAs
title_sort generation of otic lineages from integration-free human-induced pluripotent stem cells reprogrammed by mrnas
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7068143/
https://www.ncbi.nlm.nih.gov/pubmed/32190057
http://dx.doi.org/10.1155/2020/3692937
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