TY - JOUR
T1 - A Novel Fluorescent Reporter System Identifies Laminin-511/521 as Potent Regulators of Cardiomyocyte Maturation
AU - Chanthra, Nawin
AU - Abe, Tomoyuki
AU - Miyamoto, Matthew
AU - Sekiguchi, Kiyotoshi
AU - Kwon, Chulan
AU - Hanazono, Yutaka
AU - Uosaki, Hideki
N1 - Funding Information:
We gratefully acknowledge technical supports and helpful discussions from Hiromasa Hara, Suvd Byambaa, and Rie Ishihara. This work was supported by The Program for Technological Innovation of Regenerative Medicine, Research Center Network for Realization of Regenerative Medicine from Japan Agency for Medical Research and Development (AMED, 18bm0704012h0003), Grant-in-Aid for Early-Career Scientists (19K17613) and Fund for the Promotion of Joint International Research (Fostering Joint International Research (B), 19KK0219) from Japan Society for the Promotion of Science, Novartis Research Grant, Sakakibara Memorial Research Grant from Japan Research Promotion Society for Cardiovascular Diseases, Takeda Science Foundation, The Uehara Memorial Foundation, SENSHIN Medical Research Foundation, and the Grant for Basic Research of the Japanese Circulation Society (to HU). This work was also partly supported by JMU start-up award and JMU graduate student research award from Jichi Medical University (to NC), and the Projects for Technological Development, Research Center Network for Realization of Regenerative Medicine from AMED (17bm0404005h0005, to KS).
Publisher Copyright:
© 2020, The Author(s).
PY - 2020/12/1
Y1 - 2020/12/1
N2 - 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.
AB - 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.
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U2 - 10.1038/s41598-020-61163-3
DO - 10.1038/s41598-020-61163-3
M3 - Article
C2 - 32144297
AN - SCOPUS:85081531055
VL - 10
JO - Scientific Reports
JF - Scientific Reports
SN - 2045-2322
IS - 1
M1 - 4249
ER -