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Visualization of the Epiblast and Visceral Endodermal Cells Using Fgf5-P2A-Venus BAC Transgenic Mice and Epiblast Stem Cells

Fibroblast growth factor 5 (Fgf5) has been widely used as a marker for the epiblast in the postimplantation embryo and epiblast stem cells (mEpiSCs) in the mouse, making it valuable for study of differentiation of various tissues and epiblast cells in vivo and in vitro. Here, we report for the first...

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Autores principales: Khoa, Le Tran Phuc, Azami, Takuya, Tsukiyama, Tomoyuki, Matsushita, Jun, Tsukiyama-Fujii, Setsuko, Takahashi, Satoru, Ema, Masatsugu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4943650/
https://www.ncbi.nlm.nih.gov/pubmed/27409080
http://dx.doi.org/10.1371/journal.pone.0159246
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author Khoa, Le Tran Phuc
Azami, Takuya
Tsukiyama, Tomoyuki
Matsushita, Jun
Tsukiyama-Fujii, Setsuko
Takahashi, Satoru
Ema, Masatsugu
author_facet Khoa, Le Tran Phuc
Azami, Takuya
Tsukiyama, Tomoyuki
Matsushita, Jun
Tsukiyama-Fujii, Setsuko
Takahashi, Satoru
Ema, Masatsugu
author_sort Khoa, Le Tran Phuc
collection PubMed
description Fibroblast growth factor 5 (Fgf5) has been widely used as a marker for the epiblast in the postimplantation embryo and epiblast stem cells (mEpiSCs) in the mouse, making it valuable for study of differentiation of various tissues and epiblast cells in vivo and in vitro. Here, we report for the first time the generation of Fgf5-P2A-Venus BAC transgenic (Tg) mice and show that the BAC Tg can recapitulate endogenous Fgf5 expression in epiblast and visceral endodermal cells of E6.5 and 7.5 embryos. We also show that Fgf5-P2A-Venus BAC Tg mEpiSCs in the undifferentiated state expressed abundant Venus, and upon reprogramming into naïve state, Venus was suppressed. Furthermore, while most Tg mEpiSCs expressed Venus abundantly, surprisingly the Tg mEpiSCs contained a minor subpopulation of Venus-negative cells that were capable of conversion to Venus-positive cells, indicating that even Fgf5 expression shows dynamic heterogeneity in mEpiSCs. Taken together, Fgf5-P2A-Venus BAC Tg mice and mEpiSCs generated in this study will be useful for developmental biology as well as stem cell biology research.
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spelling pubmed-49436502016-08-01 Visualization of the Epiblast and Visceral Endodermal Cells Using Fgf5-P2A-Venus BAC Transgenic Mice and Epiblast Stem Cells Khoa, Le Tran Phuc Azami, Takuya Tsukiyama, Tomoyuki Matsushita, Jun Tsukiyama-Fujii, Setsuko Takahashi, Satoru Ema, Masatsugu PLoS One Research Article Fibroblast growth factor 5 (Fgf5) has been widely used as a marker for the epiblast in the postimplantation embryo and epiblast stem cells (mEpiSCs) in the mouse, making it valuable for study of differentiation of various tissues and epiblast cells in vivo and in vitro. Here, we report for the first time the generation of Fgf5-P2A-Venus BAC transgenic (Tg) mice and show that the BAC Tg can recapitulate endogenous Fgf5 expression in epiblast and visceral endodermal cells of E6.5 and 7.5 embryos. We also show that Fgf5-P2A-Venus BAC Tg mEpiSCs in the undifferentiated state expressed abundant Venus, and upon reprogramming into naïve state, Venus was suppressed. Furthermore, while most Tg mEpiSCs expressed Venus abundantly, surprisingly the Tg mEpiSCs contained a minor subpopulation of Venus-negative cells that were capable of conversion to Venus-positive cells, indicating that even Fgf5 expression shows dynamic heterogeneity in mEpiSCs. Taken together, Fgf5-P2A-Venus BAC Tg mice and mEpiSCs generated in this study will be useful for developmental biology as well as stem cell biology research. Public Library of Science 2016-07-13 /pmc/articles/PMC4943650/ /pubmed/27409080 http://dx.doi.org/10.1371/journal.pone.0159246 Text en © 2016 Khoa et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Khoa, Le Tran Phuc
Azami, Takuya
Tsukiyama, Tomoyuki
Matsushita, Jun
Tsukiyama-Fujii, Setsuko
Takahashi, Satoru
Ema, Masatsugu
Visualization of the Epiblast and Visceral Endodermal Cells Using Fgf5-P2A-Venus BAC Transgenic Mice and Epiblast Stem Cells
title Visualization of the Epiblast and Visceral Endodermal Cells Using Fgf5-P2A-Venus BAC Transgenic Mice and Epiblast Stem Cells
title_full Visualization of the Epiblast and Visceral Endodermal Cells Using Fgf5-P2A-Venus BAC Transgenic Mice and Epiblast Stem Cells
title_fullStr Visualization of the Epiblast and Visceral Endodermal Cells Using Fgf5-P2A-Venus BAC Transgenic Mice and Epiblast Stem Cells
title_full_unstemmed Visualization of the Epiblast and Visceral Endodermal Cells Using Fgf5-P2A-Venus BAC Transgenic Mice and Epiblast Stem Cells
title_short Visualization of the Epiblast and Visceral Endodermal Cells Using Fgf5-P2A-Venus BAC Transgenic Mice and Epiblast Stem Cells
title_sort visualization of the epiblast and visceral endodermal cells using fgf5-p2a-venus bac transgenic mice and epiblast stem cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4943650/
https://www.ncbi.nlm.nih.gov/pubmed/27409080
http://dx.doi.org/10.1371/journal.pone.0159246
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