{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:57:18Z","timestamp":1760147838284,"version":"3.40.5"},"reference-count":42,"publisher":"Wiley","issue":"7-8","license":[{"start":{"date-parts":[[2020,11,4]],"date-time":"2020-11-04T00:00:00Z","timestamp":1604448000000},"content-version":"vor","delay-in-days":34,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Dev Growth Differ"],"published-print":{"date-parts":[[2020,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Efficient derivation of cardiomyocytes from mouse pluripotent stem cells has proven challenging, and existing approaches rely on expensive supplementation or extensive manipulation. Mesp1 is a transcription factor that regulates cardiovascular specification during embryo development, and its overexpression has been shown to promote cardiogenesis. Here, we utilize a doxycycline\u2010inducible Mesp1\u2010expressing mouse embryonic stem cell system to develop an efficient differentiation protocol to generate functional cardiomyocytes. Our cardiac differentiation method involves transient Mesp1 induction following by subsequent dual inhibition of TGF\u03b2 and Wnt signaling pathways using small molecules. We discovered that whereas TGF\u03b2 inhibition promoted Mesp1\u2010induced cardiac differentiation, Wnt inhibition was ineffective. Nevertheless, a combined inhibition of both pathways was superior to either inhibition alone in generating cardiomyocytes. These observations suggested a potential interaction between TGF\u03b2 and Wnt signaling pathways in the context of Mesp1\u2010induced cardiac differentiation. Using a step\u2010by\u2010step approach, we have further optimized the windows of Mesp1 induction, TGF\u03b2 inhibition and Wnt inhibition to yield a maximal cardiomyocyte output \u2013 Mesp1 was induced first, followed by dual inhibition of TGF\u03b2 and Wnt signaling. Our protocol is capable of producing\u00a0approximately\u00a050% of cardiomyocytes in 12\u00a0days, which is comparable to existing methods, and have the advantages of being technically simple and inexpensive. Moreover, cardiomyocytes thus derived are functional, displaying intrinsic contractile capacity and contraction in response to electric stimulus. Derivation of mouse cardiomyocytes without the use of growth factors or other costly supplementation provides an accessible cell source for future applications.<\/jats:p>","DOI":"10.1111\/dgd.12694","type":"journal-article","created":{"date-parts":[[2020,10,13]],"date-time":"2020-10-13T19:03:44Z","timestamp":1602615824000},"page":"487-494","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Dual TGF\u03b2 and Wnt inhibition promotes Mesp1\u2010mediated mouse pluripotent stem cell differentiation into functional cardiomyocytes"],"prefix":"10.1111","volume":"62","author":[{"given":"Matthew P.","family":"Pappas","sequence":"first","affiliation":[{"name":"Department of Pediatrics University of Minnesota  Minneapolis MN USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lindsay N.","family":"Peifer","sequence":"additional","affiliation":[{"name":"Department of Pediatrics University of Minnesota  Minneapolis MN USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1432-7381","authenticated-orcid":false,"given":"Sunny S. K.","family":"Chan","sequence":"additional","affiliation":[{"name":"Department of Pediatrics University of Minnesota  Minneapolis MN USA"},{"name":"Paul and Shelia Wellstone Muscular Dystrophy Center Stem Cell Institute Lillehei Heart Institute University of Minnesota  Minneapolis MN USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2020,11,4]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0925-4773(99)00309-3"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.12703\/P5-17"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1359-6101(03)00044-3"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2008.06.009"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.2999"},{"key":"e_1_2_7_7_1","doi-asserted-by":"crossref","first-page":"876","DOI":"10.1161\/CIRCRESAHA.112.270272","article-title":"A Nodal\u2010to\u2010TGF\u03b2 cascade exerts biphasic control over cardiopoiesis","volume":"111","author":"Cai W.","year":"2011","journal-title":"Circulation Research"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2016.04.139"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.stemcr.2015.12.003"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2013.03.004"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio.137"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1582-4934.2011.01513.x"},{"key":"e_1_2_7_13_1","doi-asserted-by":"publisher","DOI":"10.1242\/dev.016865"},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12015-014-9533-0"},{"key":"e_1_2_7_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncb1696"},{"key":"e_1_2_7_16_1","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.110.221531"},{"key":"e_1_2_7_17_1","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1007\/978-1-4939-2572-8_11","article-title":"Mouse embryonic stem cell\u2010derived cardiac myocytes in a cell culture dish","volume":"1299","author":"Glass C.","year":"2015","journal-title":"Methods in Molecular Biology"},{"key":"e_1_2_7_18_1","doi-asserted-by":"publisher","DOI":"10.1038\/cr.2008.302"},{"key":"e_1_2_7_19_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1120299109"},{"key":"e_1_2_7_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2010.12.008"},{"key":"e_1_2_7_21_1","doi-asserted-by":"publisher","DOI":"10.1242\/dev.127.15.3215"},{"key":"e_1_2_7_22_1","doi-asserted-by":"publisher","DOI":"10.1002\/dvdy.24366"},{"key":"e_1_2_7_23_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.aao4174"},{"key":"e_1_2_7_24_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.3448"},{"key":"e_1_2_7_25_1","doi-asserted-by":"publisher","DOI":"10.1242\/dev.02551"},{"key":"e_1_2_7_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2008.04.004"},{"key":"e_1_2_7_27_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0609100104"},{"key":"e_1_2_7_28_1","article-title":"Mesp1 marked cardiac progenitor cells repair infarcted mouse hearts","volume":"6","author":"Liu Y. 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