A Novel Fluorescent Reporter System Identifies Laminin-511/521 as Potent Regulators of Cardiomyocyte Maturation

Pluripotent stem cell-derived cardiomyocytes (PSC-CMs) hold great promise for disease modeling and drug discovery. However, PSC-CMs exhibit immature phenotypes in culture, and the lack of maturity limits their broad applications. While physical and functional analyses are generally used to determine...

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Veröffentlicht in:	Scientific reports 2020-03, Vol.10 (1), p.4249-4249, Article 4249
Hauptverfasser:	Chanthra, Nawin, Abe, Tomoyuki, Miyamoto, Matthew, Sekiguchi, Kiyotoshi, Kwon, Chulan, Hanazono, Yutaka, Uosaki, Hideki
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Sprache:	eng
Schlagworte:	631/136 631/532/2064 631/61/32 631/80/79/750 692/4019/592 Biomarkers Calcium Cardiomyocytes Cell culture Cell Differentiation - genetics Computational Biology - methods Cytology Embryo cells Enhancers Extracellular Matrix - metabolism Gene Expression Gene Expression Profiling Genes, Reporter Humanities and Social Sciences Laminin Laminin - genetics Laminin - metabolism Maturation multidisciplinary Myocytes, Cardiac - cytology Myocytes, Cardiac - metabolism Phenotypes Pluripotency Pluripotent Stem Cells - cytology Pluripotent Stem Cells - metabolism Sarcomeres - metabolism Science Science (multidisciplinary) Stem cell transplantation Stem cells Transcription Transcription, Genetic Transcriptome
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creator	Chanthra, Nawin Abe, Tomoyuki Miyamoto, Matthew Sekiguchi, Kiyotoshi Kwon, Chulan Hanazono, Yutaka Uosaki, Hideki
description	Pluripotent stem cell-derived cardiomyocytes (PSC-CMs) hold great promise for disease modeling and drug discovery. However, PSC-CMs exhibit immature phenotypes in culture, and the lack of maturity limits their broad applications. While physical and functional analyses are generally used to determine the status of cardiomyocyte maturation, they could be time-consuming and often present challenges in comparing maturation-enhancing strategies. Therefore, there is a demand for a method to assess cardiomyocyte maturation rapidly and reproducibly. In this study, we found that Myomesin-2 ( Myom2 ), encoding M-protein, is upregulated postnatally, and based on this, we targeted TagRFP to the Myom2 locus in mouse embryonic stem cells. Myom2-RFP + PSC-CMs exhibited more mature phenotypes than RFP - cells in morphology, function and transcriptionally, conductive to sarcomere shortening assays. Using this system, we screened extracellular matrices (ECMs) and identified laminin-511/521 as potent enhancers of cardiomyocyte maturation. Together, we developed and characterized a novel fluorescent reporter system for the assessment of cardiomyocyte maturation and identified potent maturation-enhancing ECMs through this simple and rapid assay. This system is expected to facilitate use of PSC-CMs in a variety of scientific and medical investigations.
doi_str_mv	10.1038/s41598-020-61163-3
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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Scientific reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chanthra, Nawin</au><au>Abe, Tomoyuki</au><au>Miyamoto, Matthew</au><au>Sekiguchi, Kiyotoshi</au><au>Kwon, Chulan</au><au>Hanazono, Yutaka</au><au>Uosaki, Hideki</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A Novel Fluorescent Reporter System Identifies Laminin-511/521 as Potent Regulators of Cardiomyocyte Maturation</atitle><jtitle>Scientific reports</jtitle><stitle>Sci Rep</stitle><addtitle>Sci Rep</addtitle><date>2020-03-06</date><risdate>2020</risdate><volume>10</volume><issue>1</issue><spage>4249</spage><epage>4249</epage><pages>4249-4249</pages><artnum>4249</artnum><issn>2045-2322</issn><eissn>2045-2322</eissn><abstract>Pluripotent stem cell-derived cardiomyocytes (PSC-CMs) hold great promise for disease modeling and drug discovery. However, PSC-CMs exhibit immature phenotypes in culture, and the lack of maturity limits their broad applications. While physical and functional analyses are generally used to determine the status of cardiomyocyte maturation, they could be time-consuming and often present challenges in comparing maturation-enhancing strategies. Therefore, there is a demand for a method to assess cardiomyocyte maturation rapidly and reproducibly. In this study, we found that Myomesin-2 ( Myom2 ), encoding M-protein, is upregulated postnatally, and based on this, we targeted TagRFP to the Myom2 locus in mouse embryonic stem cells. Myom2-RFP + PSC-CMs exhibited more mature phenotypes than RFP - cells in morphology, function and transcriptionally, conductive to sarcomere shortening assays. Using this system, we screened extracellular matrices (ECMs) and identified laminin-511/521 as potent enhancers of cardiomyocyte maturation. Together, we developed and characterized a novel fluorescent reporter system for the assessment of cardiomyocyte maturation and identified potent maturation-enhancing ECMs through this simple and rapid assay. This system is expected to facilitate use of PSC-CMs in a variety of scientific and medical investigations.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>32144297</pmid><doi>10.1038/s41598-020-61163-3</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record>
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subjects	631/136 631/532/2064 631/61/32 631/80/79/750 692/4019/592 Biomarkers Calcium Cardiomyocytes Cell culture Cell Differentiation - genetics Computational Biology - methods Cytology Embryo cells Enhancers Extracellular Matrix - metabolism Gene Expression Gene Expression Profiling Genes, Reporter Humanities and Social Sciences Laminin Laminin - genetics Laminin - metabolism Maturation multidisciplinary Myocytes, Cardiac - cytology Myocytes, Cardiac - metabolism Phenotypes Pluripotency Pluripotent Stem Cells - cytology Pluripotent Stem Cells - metabolism Sarcomeres - metabolism Science Science (multidisciplinary) Stem cell transplantation Stem cells Transcription Transcription, Genetic Transcriptome
title	A Novel Fluorescent Reporter System Identifies Laminin-511/521 as Potent Regulators of Cardiomyocyte Maturation
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