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Recapitulating kidney development in vitro by priming and differentiating mouse embryonic stem cells in monolayers

In order to harness the potential of pluripotent stem cells, we need to understand how to differentiate them to our target cell types. Here, we developed a protocol to differentiate mouse embryonic stem cells (ESCs) to renal progenitors in a step-wise manner. Microarrays were used to track the trans...

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Autores principales: Chow, Theresa, Wong, Frances T. M., Monetti, Claudio, Nagy, Andras, Cox, Brian, Rogers, Ian M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7171095/
https://www.ncbi.nlm.nih.gov/pubmed/32351711
http://dx.doi.org/10.1038/s41536-020-0092-5
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author Chow, Theresa
Wong, Frances T. M.
Monetti, Claudio
Nagy, Andras
Cox, Brian
Rogers, Ian M.
author_facet Chow, Theresa
Wong, Frances T. M.
Monetti, Claudio
Nagy, Andras
Cox, Brian
Rogers, Ian M.
author_sort Chow, Theresa
collection PubMed
description In order to harness the potential of pluripotent stem cells, we need to understand how to differentiate them to our target cell types. Here, we developed a protocol to differentiate mouse embryonic stem cells (ESCs) to renal progenitors in a step-wise manner. Microarrays were used to track the transcriptional changes at each stage of differentiation and we observed that genes associated with metanephros, ureteric bud, and blood vessel development were significantly upregulated as the cells differentiated towards renal progenitors. Priming the ESCs and optimizing seeding cell density and growth factor concentrations helped improve differentiation efficiency. Organoids were used to determine the developmental potential of the renal progenitor cells. Aggregated renal progenitors gave rise to organoids consisting of LTL+/E-cadherin+ proximal tubules, cytokeratin+ ureteric bud-derived tubules, and extracellular matrix proteins secreted by the cells themselves. Over-expression of key kidney developmental genes, Pax2, Six1, Eya1, and Hox11 paralogs, during differentiation did not improve differentiation efficiency. Altogether, we developed a protocol to differentiate mouse ESCs in a manner that recapitulates embryonic kidney development and showed that precise gene regulation is essential for proper differentiation to occur.
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spelling pubmed-71710952020-04-29 Recapitulating kidney development in vitro by priming and differentiating mouse embryonic stem cells in monolayers Chow, Theresa Wong, Frances T. M. Monetti, Claudio Nagy, Andras Cox, Brian Rogers, Ian M. NPJ Regen Med Article In order to harness the potential of pluripotent stem cells, we need to understand how to differentiate them to our target cell types. Here, we developed a protocol to differentiate mouse embryonic stem cells (ESCs) to renal progenitors in a step-wise manner. Microarrays were used to track the transcriptional changes at each stage of differentiation and we observed that genes associated with metanephros, ureteric bud, and blood vessel development were significantly upregulated as the cells differentiated towards renal progenitors. Priming the ESCs and optimizing seeding cell density and growth factor concentrations helped improve differentiation efficiency. Organoids were used to determine the developmental potential of the renal progenitor cells. Aggregated renal progenitors gave rise to organoids consisting of LTL+/E-cadherin+ proximal tubules, cytokeratin+ ureteric bud-derived tubules, and extracellular matrix proteins secreted by the cells themselves. Over-expression of key kidney developmental genes, Pax2, Six1, Eya1, and Hox11 paralogs, during differentiation did not improve differentiation efficiency. Altogether, we developed a protocol to differentiate mouse ESCs in a manner that recapitulates embryonic kidney development and showed that precise gene regulation is essential for proper differentiation to occur. Nature Publishing Group UK 2020-04-20 /pmc/articles/PMC7171095/ /pubmed/32351711 http://dx.doi.org/10.1038/s41536-020-0092-5 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Chow, Theresa
Wong, Frances T. M.
Monetti, Claudio
Nagy, Andras
Cox, Brian
Rogers, Ian M.
Recapitulating kidney development in vitro by priming and differentiating mouse embryonic stem cells in monolayers
title Recapitulating kidney development in vitro by priming and differentiating mouse embryonic stem cells in monolayers
title_full Recapitulating kidney development in vitro by priming and differentiating mouse embryonic stem cells in monolayers
title_fullStr Recapitulating kidney development in vitro by priming and differentiating mouse embryonic stem cells in monolayers
title_full_unstemmed Recapitulating kidney development in vitro by priming and differentiating mouse embryonic stem cells in monolayers
title_short Recapitulating kidney development in vitro by priming and differentiating mouse embryonic stem cells in monolayers
title_sort recapitulating kidney development in vitro by priming and differentiating mouse embryonic stem cells in monolayers
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7171095/
https://www.ncbi.nlm.nih.gov/pubmed/32351711
http://dx.doi.org/10.1038/s41536-020-0092-5
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