title: Functional cardiomyocytes derived from Isl1 cardiac progenitors via Bmp4 stimulation.
abstract: As heart failure due to myocardial infarction remains a leading cause of morbidity worldwide, cell-based cardiac regenerative therapy using cardiac progenitor cells (CPCs) could provide a potential treatment for the repair of injured myocardium. As adult CPCs may have limitations regarding tissue accessibility and proliferative ability, CPCs derived from embryonic stem cells (ESCs) could serve as an unlimited source of cells with high proliferative ability. As one of the CPCs that can be derived from embryonic stem cells, Isl1 expressing cardiac progenitor cells (Isl1-CPCs) may serve as a valuable source of cells for cardiac repair due to their high cardiac differentiation potential and authentic cardiac origin. In order to generate an unlimited number of Isl1-CPCs, we used a previously established an ESC line that allows for isolation of Isl1-CPCs by green fluorescent protein (GFP) expression that is directed by the mef2c gene, specifically expressed in the Isl1 domain of the anterior heart field. To improve the efficiency of cardiac differentiation of Isl1-CPCs, we studied the role of Bmp4 in cardiogenesis of Isl1-CPCs. We show an inductive role of Bmp directly on cardiac progenitors and its enhancement on early cardiac differentiation of CPCs. Upon induction of Bmp4 to Isl1-CPCs during differentiation, the cTnT+ cardiomyocyte population was enhanced 2.8±0.4 fold for Bmp4 treated CPC cultures compared to that detected for vehicle treated cultures. Both Bmp4 treated and untreated cardiomyocytes exhibit proper electrophysiological and calcium signaling properties. In addition, we observed a significant increase in Tbx5 and Tbx20 expression in differentiation cultures treated with Bmp4 compared to the untreated control, suggesting a link between Bmp4 and Tbx genes which may contribute to the enhanced cardiac differentiation in Bmp4 treated cultures. Collectively these findings suggest a cardiomyogenic role for Bmp4 directly on a pure population of Isl1 expressing cardiac progenitors, which could lead to enhancement of cardiac differentiation and engraftment, holding a significant therapeutic value for cardiac repair in the future.
authors.affiliation: Yale Cardiovascular Research Center, Section of Cardiovascular Medicine, Department of Internal Medicine, Yale Stem Cell Center, Yale School of Medicine, Yale University, New Haven, CT, United States of America; Department of Medical Biology, School of Medicine, Istanbul Medipol University, Istanbul, Turkey.
authors.forename: Esra
authors.initials: E
authors.lastname: Cagavi
authors.affiliation: Yale Cardiovascular Research Center, Section of Cardiovascular Medicine, Department of Internal Medicine, Yale Stem Cell Center, Yale School of Medicine, Yale University, New Haven, CT, United States of America.
authors.forename: Oscar
authors.initials: O
authors.lastname: Bartulos
authors.affiliation: Yale Cardiovascular Research Center, Section of Cardiovascular Medicine, Department of Internal Medicine, Yale Stem Cell Center, Yale School of Medicine, Yale University, New Haven, CT, United States of America; Department of Genetics, Yale School of Medicine, Yale University, New Haven, CT, United States of America.
authors.forename: Carol Y
authors.initials: CY
authors.lastname: Suh
authors.affiliation: Department of Cell Biology, University of Connecticut Health Center, Farmington, CT, United States of America.
authors.forename: Baonan
authors.initials: B
authors.lastname: Sun
authors.affiliation: Department of Cell Biology, University of Connecticut Health Center, Farmington, CT, United States of America.
authors.forename: Zhichao
authors.initials: Z
authors.lastname: Yue
authors.affiliation: Yale Cardiovascular Research Center, Section of Cardiovascular Medicine, Department of Internal Medicine, Yale Stem Cell Center, Yale School of Medicine, Yale University, New Haven, CT, United States of America.
authors.forename: Zhengxin
authors.initials: Z
authors.lastname: Jiang
authors.affiliation: Department of Cell Biology, University of Connecticut Health Center, Farmington, CT, United States of America.
authors.forename: Lixia
authors.initials: L
authors.lastname: Yue
authors.affiliation: Yale Cardiovascular Research Center, Section of Cardiovascular Medicine, Department of Internal Medicine, Yale Stem Cell Center, Yale School of Medicine, Yale University, New Haven, CT, United States of America; Department of Pathology, Yale School of Medicine, New Haven, CT, United States of America; Vascular Biology and Therapeutics Program, Yale University School of Medicine, New Haven, CT, United States of America.
authors.forename: Yibing
authors.initials: Y
authors.lastname: Qyang
mesh_terms: D000200:Action Potentials; D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology*; D020013:Calcium Signaling; D002454:Cell Differentiation; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D060850:LIM-Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D020825:T-Box Domain Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D055415:Bone Morphogenetic Protein 4; D060850:LIM-Homeodomain Proteins; D020825:T-Box Domain Proteins; C103172:T-box transcription factor 5; C486013:Tbx20 protein, mouse; D014157:Transcription Factors; C063843:insulin gene enhancer binding protein Isl-1