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Overexpression of GATA binding protein 4 and myocyte enhancer factor 2C induces differentiation of mesenchymal stem cells into cardiac-like cells

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World Journal of Stem Cells 2022年第9期14卷 700-713页

作者： Syeda Saima Razzaq Irfan Khan Nadia Naeem Asmat Salim Sumreen Begum kanwal haneef University of Karachi Karachi 75270Pakistan Panjwani Center for Molecular Medicine and Drug Research International Center for Chemical and Biological SciencesUniversity of KarachiKarachi 75270Pakistan Dow Research Institute of Biotechnology&Biomedical Sciences(DRIBBS) Dow University of Health Sciences(DUHS)Ojha CampusKarachi 75200Pakistan Stem Cells Research Laboratory(SCRL) Sindh Institute of Urology and Transplantation(SIUT)Karachi 74200Pakistan

BACKGROUND Heart diseases are the primary cause of death all over the world.Following myocardial infarction,billions of cells die,resulting in a huge loss of cardiac function.Stem cell-based therapies have appeared as a new area to support heart regeneration.The transcription factors GATA binding protein 4(GATA-4)and myocyte enhancer factor 2C(MEF2C)are considered prominent factors in the development of the cardiovascular system.AIM To explore the potential of GATA-4 and MEF2C for the cardiac differentiation of human umbilical cord mesenchymal stem cells(hUC-MSCs).METHODS hUC-MSCs were characterized morphologically and immunologically by the presence of specific markers of MSCs via immunocytochemistry and flow cytometry,and by their potential to differentiate into osteocytes and adipocytes.hUC-MSCs were transfected with GATA-4,MEF2C,and their combination to direct the differentiation.Cardiac differentiation was confirmed by semiquant itative real-time polymerase chain reaction and immunocytochemistry.RESULTS hUC-MSCs expressed specific cell surface markers CD105,CD90,CD44,and vimentin but lack the expression of CD45.The transcription factors GATA-4 and MEF2C,and their combination induced differentiation in hUC-MSCs with significant expression of cardiac genes i.e.,GATA-4,MEF2C,NK2 homeobox 5(NKX2.5),MHC,and connexin-43,and cardiac proteins GATA-4,NKX2.5,cardiac troponin T,and connexin-43.CONCLUSION Transfection with GATA-4,MEF2C,and their combination effectively induces cardiac differentiation in hUC-MSCs.These genetically modified MSCs could be a promising treatment option for heart diseases in the future.

关键词： Heart disease GATA binding protein 4 Myocyte enhancer factor 2C Transcription factors Differentiation Human umbilical cord-mesenchymal stem cells

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Combination of mesenchymal stem cells and three-dimensional collagen scaffold preserves ventricular remodeling in rat myocardial infarction model

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World Journal of Stem Cells 2022年第8期14卷 633-657页

作者： Rida-e-Maria Qazi Irfan Khan kanwal haneef Tuba Shakil Malick Nadia Naeem Waqas Ahmad Asmat Salim Sadia Mohsin Stem Cell Laboratory Dr.Panjwani Center for Molecular Medicine and Drug ResearchICCBSUniversity of KarachiKarachi 75270SindhPakistan Dr.Zafar H.Zaidi Center for Proteomics University of KarachiKarachi 75270SindhPakistan Dow Research Institute of Biotechnology and Biomedical Sciences(DRIBBS) Dow University of Health and SciencesOjha CampusKarachi 74200SindhPakistan Cardiovascular Research Center Lewis Katz School of MedicineTemple UniversityPhiladelphiaPA 19140United States

BACKGROUND Cardiovascular diseases are the major cause of mortality worldwide.Regeneration of the damaged myocardium remains a challenge due to mechanical constraints and limited healing ability of the adult heart tissue.Cardiac tissue engineering using biomaterial scaffolds combined with stem cells and bioactive molecules could be a highly promising approach for cardiac repair.Use of biomaterials can provide suitable microenvironment to the cells and can solve cell engraftment problems associated with cell transplantation alone.Mesenchymal stem cells(MSCs)are potential candidates in cardiac tissue engineering because of their multilineage differentiation potential and ease of isolation.Use of DNA methyl transferase inhibitor,such as zebularine,in combination with three-dimensional(3D)scaffold can promote efficient MSC differentiation into cardiac lineage,as epigenetic modifications play a fundamental role in determining cell fate and lineage specific gene expression.AIM To investigate the role of collagen scaffold and zebularine in the differentiation of rat bone marrow(BM)-MSCs and their subsequent in vivo effects.METHODS MSCs were isolated from rat BM and characterized morphologically,immunophenotypically and by multilineage differentiation potential.MSCs were seeded in collagen scaffold and treated with 3μmol/L zebularine in three different ways.Cytotoxicity analysis was done and cardiac differentiation was analyzed at the gene and protein levels.Treated and untreated MSC-seeded scaffolds were transplanted in the rat myocardial infarction(MI)model and cardiac function was assessed by echocardiography.Cell tracking was performed by DiI dye labeling,while regeneration and neovascularization were evaluated by histological and immunohistochemical analysis,respectively.RESULTS MSCs were successfully isolated and seeded in collagen scaffold.Cytotoxicity analysis revealed that zebularine was not cytotoxic in any of the treatment groups.Cardiac differentiation analysis showed more pronounced results in the type 3 tre

关键词： Mesenchymal stem cells Myocardial infarction Cardiac tissue engineering Demethylating agent Collagen scaffold Zebularine

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