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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2020
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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. |
format | Online Article Text |
id | pubmed-7171095 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
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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