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Activation of Wnt/ß-catenin signaling in ESC promotes rostral forebrain differentiation in vitro

Wnt/ß-catenin signaling is crucial for maintenance of pluripotent state of embryonic stem cell (ESC). However, it is unclear how Wnt/ß-catenin signaling affects the differentiation ability of ESC, especially with regard to rostral forebrain cells. Here, using Rax, rostral forebrain marker, and Wnt/ß...

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Autores principales: Takata, Nozomu, Sakakura, Eriko, Sasai, Yoshiki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer US 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4833820/
https://www.ncbi.nlm.nih.gov/pubmed/26563247
http://dx.doi.org/10.1007/s11626-015-9975-y
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author Takata, Nozomu
Sakakura, Eriko
Sasai, Yoshiki
author_facet Takata, Nozomu
Sakakura, Eriko
Sasai, Yoshiki
author_sort Takata, Nozomu
collection PubMed
description Wnt/ß-catenin signaling is crucial for maintenance of pluripotent state of embryonic stem cell (ESC). However, it is unclear how Wnt/ß-catenin signaling affects the differentiation ability of ESC, especially with regard to rostral forebrain cells. Here, using Rax, rostral forebrain marker, and Wnt/ß-catenin reporter lines, we report ratio of Rax(+) and Wnt responding tissue (Wnt(+)) patterns, which were affected by seeding number of ESC in three-dimensional culture system. Surprisingly, we found ß-catenin level and localization are heterogeneous in ESC colony by immunostaining and time-laps imaging of ß-catenin-mEGFP signals. Moreover, activation of Wnt signaling in ESC promoted expression level and nuclear localization of ß-catenin, and mRNA levels of Wnt antagonists, axin2 and dkk1, leading to upregulating Wnt/ß-catenin reporter in ESC state and Rax expression at differentiation culture day 7. Together, our results suggest that activation of Wnt signaling in ESC promotes the differentiation efficacy of rostral forebrain cells. Wnt-priming culture method may provide a useful tool for applications in the areas of basic science and molecular therapeutics for regenerative medicine. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11626-015-9975-y) contains supplementary material, which is available to authorized users.
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spelling pubmed-48338202016-04-25 Activation of Wnt/ß-catenin signaling in ESC promotes rostral forebrain differentiation in vitro Takata, Nozomu Sakakura, Eriko Sasai, Yoshiki In Vitro Cell Dev Biol Anim Article Wnt/ß-catenin signaling is crucial for maintenance of pluripotent state of embryonic stem cell (ESC). However, it is unclear how Wnt/ß-catenin signaling affects the differentiation ability of ESC, especially with regard to rostral forebrain cells. Here, using Rax, rostral forebrain marker, and Wnt/ß-catenin reporter lines, we report ratio of Rax(+) and Wnt responding tissue (Wnt(+)) patterns, which were affected by seeding number of ESC in three-dimensional culture system. Surprisingly, we found ß-catenin level and localization are heterogeneous in ESC colony by immunostaining and time-laps imaging of ß-catenin-mEGFP signals. Moreover, activation of Wnt signaling in ESC promoted expression level and nuclear localization of ß-catenin, and mRNA levels of Wnt antagonists, axin2 and dkk1, leading to upregulating Wnt/ß-catenin reporter in ESC state and Rax expression at differentiation culture day 7. Together, our results suggest that activation of Wnt signaling in ESC promotes the differentiation efficacy of rostral forebrain cells. Wnt-priming culture method may provide a useful tool for applications in the areas of basic science and molecular therapeutics for regenerative medicine. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11626-015-9975-y) contains supplementary material, which is available to authorized users. Springer US 2015-11-12 2016 /pmc/articles/PMC4833820/ /pubmed/26563247 http://dx.doi.org/10.1007/s11626-015-9975-y Text en © The Author(s) 2015 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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.
spellingShingle Article
Takata, Nozomu
Sakakura, Eriko
Sasai, Yoshiki
Activation of Wnt/ß-catenin signaling in ESC promotes rostral forebrain differentiation in vitro
title Activation of Wnt/ß-catenin signaling in ESC promotes rostral forebrain differentiation in vitro
title_full Activation of Wnt/ß-catenin signaling in ESC promotes rostral forebrain differentiation in vitro
title_fullStr Activation of Wnt/ß-catenin signaling in ESC promotes rostral forebrain differentiation in vitro
title_full_unstemmed Activation of Wnt/ß-catenin signaling in ESC promotes rostral forebrain differentiation in vitro
title_short Activation of Wnt/ß-catenin signaling in ESC promotes rostral forebrain differentiation in vitro
title_sort activation of wnt/ß-catenin signaling in esc promotes rostral forebrain differentiation in vitro
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4833820/
https://www.ncbi.nlm.nih.gov/pubmed/26563247
http://dx.doi.org/10.1007/s11626-015-9975-y
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