Specialized mouse embryonic stem cells for studying vascular development

Drew E Glaser,1 Andrew B Burns,2 Rachel Hatano,2 Magdalena Medrzycki,3 Yuhong Fan,3 Kara E McCloskey1 1School of Engineering, University of California, Merced, CA, USA; 2School of Natural Sciences, University of California, Merced, CA, USA; 3School of Biology and the Petit Institute for Bioengineer...

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Autores principales:	Glaser DE, Burns AB, Hatano R, Medrzycki M, Fan Y, McCloskey KE
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2014
Materias:	Cytology QH573-671
Acceso en línea:	https://doaj.org/article/ebf3d6425e684bbfb57a2b26f5e94150
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spelling	oai:doaj.org-article:ebf3d6425e684bbfb57a2b26f5e941502021-12-02T06:27:45ZSpecialized mouse embryonic stem cells for studying vascular development1178-6957https://doaj.org/article/ebf3d6425e684bbfb57a2b26f5e941502014-10-01T00:00:00Zhttp://www.dovepress.com/specialized-mouse-embryonic-stem-cells-for-studying-vascular-developme-peer-reviewed-article-SCCAAhttps://doaj.org/toc/1178-6957 Drew E Glaser,1 Andrew B Burns,2 Rachel Hatano,2 Magdalena Medrzycki,3 Yuhong Fan,3 Kara E McCloskey1 1School of Engineering, University of California, Merced, CA, USA; 2School of Natural Sciences, University of California, Merced, CA, USA; 3School of Biology and the Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA, USAAbstract: Vascular progenitor cells are desirable in a variety of therapeutic strategies; however, the lineage commitment of endothelial and smooth muscle cell from a common progenitor is not well-understood. Here, we report the generation of the first dual reporter mouse embryonic stem cell (mESC) lines designed to facilitate the study of vascular endothelial and smooth muscle development in vitro. These mESC lines express green fluorescent protein (GFP) under the endothelial promoter, Tie-2, and Discomsoma sp. red fluorescent protein (RFP) under the promoter for alpha-smooth muscle actin (α-SMA). The lines were then characterized for morphology, marker expression, and pluripotency. The mESC colonies were found to exhibit dome-shaped morphology, alkaline phosphotase activity, as well as expression of Oct 3/4 and stage-specific embryonic antigen-1. The mESC colonies were also found to display normal karyotypes and are able to generate cells from all three germ layers, verifying pluripotency. Tissue staining confirmed the coexpression of VE (vascular endothelial)-cadherin with the Tie-2 GFP+ expression on endothelial structures and smooth muscle myosin heavy chain with the α-SMA RFP+ smooth muscle cells. Lastly, it was verified that the developing mESC do express Tie-2 GFP+ and α-SMA RFP+ cells during differentiation and that the GFP+ cells colocalize with the vascular-like structures surrounded by α-SMA-RFP cells. These dual reporter vascular-specific mESC permit visualization and cell tracking of individual endothelial and smooth muscle cells over time and in multiple dimensions, a powerful new tool for studying vascular development in real time.Keywords: vascular progenitor cells, endothelial cells, smooth muscle cells, embryoid body, vasculogenesisGlaser DEBurns ABHatano RMedrzycki MFan YMcCloskey KEDove Medical PressarticleCytologyQH573-671ENStem Cells and Cloning: Advances and Applications, Vol 2014, Iss default, Pp 79-88 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Cytology QH573-671
spellingShingle	Cytology QH573-671 Glaser DE Burns AB Hatano R Medrzycki M Fan Y McCloskey KE Specialized mouse embryonic stem cells for studying vascular development
description	Drew E Glaser,1 Andrew B Burns,2 Rachel Hatano,2 Magdalena Medrzycki,3 Yuhong Fan,3 Kara E McCloskey1 1School of Engineering, University of California, Merced, CA, USA; 2School of Natural Sciences, University of California, Merced, CA, USA; 3School of Biology and the Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA, USAAbstract: Vascular progenitor cells are desirable in a variety of therapeutic strategies; however, the lineage commitment of endothelial and smooth muscle cell from a common progenitor is not well-understood. Here, we report the generation of the first dual reporter mouse embryonic stem cell (mESC) lines designed to facilitate the study of vascular endothelial and smooth muscle development in vitro. These mESC lines express green fluorescent protein (GFP) under the endothelial promoter, Tie-2, and Discomsoma sp. red fluorescent protein (RFP) under the promoter for alpha-smooth muscle actin (α-SMA). The lines were then characterized for morphology, marker expression, and pluripotency. The mESC colonies were found to exhibit dome-shaped morphology, alkaline phosphotase activity, as well as expression of Oct 3/4 and stage-specific embryonic antigen-1. The mESC colonies were also found to display normal karyotypes and are able to generate cells from all three germ layers, verifying pluripotency. Tissue staining confirmed the coexpression of VE (vascular endothelial)-cadherin with the Tie-2 GFP+ expression on endothelial structures and smooth muscle myosin heavy chain with the α-SMA RFP+ smooth muscle cells. Lastly, it was verified that the developing mESC do express Tie-2 GFP+ and α-SMA RFP+ cells during differentiation and that the GFP+ cells colocalize with the vascular-like structures surrounded by α-SMA-RFP cells. These dual reporter vascular-specific mESC permit visualization and cell tracking of individual endothelial and smooth muscle cells over time and in multiple dimensions, a powerful new tool for studying vascular development in real time.Keywords: vascular progenitor cells, endothelial cells, smooth muscle cells, embryoid body, vasculogenesis
format	article
author	Glaser DE Burns AB Hatano R Medrzycki M Fan Y McCloskey KE
author_facet	Glaser DE Burns AB Hatano R Medrzycki M Fan Y McCloskey KE
author_sort	Glaser DE
title	Specialized mouse embryonic stem cells for studying vascular development
title_short	Specialized mouse embryonic stem cells for studying vascular development
title_full	Specialized mouse embryonic stem cells for studying vascular development
title_fullStr	Specialized mouse embryonic stem cells for studying vascular development
title_full_unstemmed	Specialized mouse embryonic stem cells for studying vascular development
title_sort	specialized mouse embryonic stem cells for studying vascular development
publisher	Dove Medical Press
publishDate	2014
url	https://doaj.org/article/ebf3d6425e684bbfb57a2b26f5e94150
work_keys_str_mv	AT glaserde specializedmouseembryonicstemcellsforstudyingvasculardevelopment AT burnsab specializedmouseembryonicstemcellsforstudyingvasculardevelopment AT hatanor specializedmouseembryonicstemcellsforstudyingvasculardevelopment AT medrzyckim specializedmouseembryonicstemcellsforstudyingvasculardevelopment AT fany specializedmouseembryonicstemcellsforstudyingvasculardevelopment AT mccloskeyke specializedmouseembryonicstemcellsforstudyingvasculardevelopment
_version_	1718399908780703744

Specialized mouse embryonic stem cells for studying vascular development

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