title: Wnt/β-Catenin Stimulation and Laminins Support Cardiovascular Cell Progenitor Expansion from Human Fetal Cardiac Mesenchymal Stromal Cells.
abstract: The intrinsic regenerative capacity of human fetal cardiac mesenchymal stromal cells (MSCs) has not been fully characterized. Here we demonstrate that we can expand cells with characteristics of cardiovascular progenitor cells from the MSC population of human fetal hearts. Cells cultured on cardiac muscle laminin (LN)-based substrata in combination with stimulation of the canonical Wnt/β-catenin pathway showed increased gene expression of ISL1, OCT4, KDR, and NKX2.5. The majority of cells stained positive for PDGFR-α, ISL1, and NKX2.5, and subpopulations also expressed the progenitor markers TBX18, KDR, c-KIT, and SSEA-1. Upon culture of the cardiac MSCs in differentiation media and on relevant LNs, portions of the cells differentiated into spontaneously beating cardiomyocytes, and endothelial and smooth muscle-like cells. Our protocol for large-scale culture of human fetal cardiac MSCs enables future exploration of the regenerative functions of these cells in the context of myocardial injury in vitro and in vivo.
authors.affiliation: Division of Cardiology, Department of Medicine, Karolinska Institutet, 17177 Stockholm, Sweden.
authors.forename: Agneta
authors.initials: A
authors.lastname: Månsson-Broberg
authors.affiliation: Division of Matrix Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 17177 Stockholm, Sweden.
authors.forename: Sergey
authors.initials: S
authors.lastname: Rodin
authors.affiliation: Division of Cardiothoracic Surgery and Anesthesiology, Department of Molecular Medicine and Surgery, Karolinska Institutet, 17177 Stockholm, Sweden.
authors.forename: Ivana
authors.initials: I
authors.lastname: Bulatovic
authors.affiliation: Division of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 17177 Stockholm, Sweden; Cardiovascular & Metabolic Diseases, Innovative Medicines and Early Development, AstraZeneca R&D, 43150 Mölndal, Sweden.
authors.forename: Cristián
authors.initials: C
authors.lastname: Ibarra
authors.affiliation: Division of Cardiothoracic Surgery and Anesthesiology, Department of Molecular Medicine and Surgery, Karolinska Institutet, 17177 Stockholm, Sweden.
authors.forename: Marie
authors.initials: M
authors.lastname: Löfling
authors.affiliation: Division of Cardiothoracic Surgery and Anesthesiology, Department of Molecular Medicine and Surgery, Karolinska Institutet, 17177 Stockholm, Sweden.
authors.forename: Rami
authors.initials: R
authors.lastname: Genead
authors.affiliation: Division of Cardiology, Department of Medicine, Karolinska Institutet, 17177 Stockholm, Sweden.
authors.forename: Eva
authors.initials: E
authors.lastname: Wärdell
authors.affiliation: Division of Cardiothoracic Surgery and Anesthesiology, Department of Molecular Medicine and Surgery, Karolinska Institutet, 17177 Stockholm, Sweden.
authors.forename: Ulrika
authors.initials: U
authors.lastname: Felldin
authors.affiliation: Division of Cardiothoracic Surgery and Anesthesiology, Department of Molecular Medicine and Surgery, Karolinska Institutet, 17177 Stockholm, Sweden.
authors.forename: Carl
authors.initials: C
authors.lastname: Granath
authors.affiliation: Division of Hematology, Department of Medicine, Karolinska Institutet, Karolinska University Hospital, 17177 Stockholm, Sweden; Cell Therapy Institute, Nova Southeastern University, Fort Lauderdale, FL 33314, USA.
authors.forename: Evren
authors.initials: E
authors.lastname: Alici
authors.affiliation: Division of Hematology, Department of Medicine, Karolinska Institutet, Karolinska University Hospital, 17177 Stockholm, Sweden; Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital, 17177 Stockholm, Sweden.
authors.forename: Katarina
authors.initials: K
authors.lastname: Le Blanc
authors.affiliation: Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital, 17177 Stockholm, Sweden.
authors.forename: C I Edvard
authors.initials: CIE
authors.lastname: Smith
authors.affiliation: Division of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 17177 Stockholm, Sweden.
authors.forename: Alena
authors.initials: A
authors.lastname: Salašová
authors.affiliation: CLINTEC, Division of Obstetrics and Gynecology, Karolinska Institutet, Karolinska University Hospital, 17177 Stockholm, Sweden.
authors.forename: Magnus
authors.initials: M
authors.lastname: Westgren
authors.affiliation: Division of Neurodegeneration, Department of Neurobiology, Care Sciences and Society, Karolinska Institutet, Karolinska University Hospital, 17177 Stockholm, Sweden.
authors.forename: Erik
authors.initials: E
authors.lastname: Sundström
authors.affiliation: Division of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 17177 Stockholm, Sweden.
authors.forename: Per
authors.initials: P
authors.lastname: Uhlén
authors.affiliation: Division of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 17177 Stockholm, Sweden.
authors.forename: Ernest
authors.initials: E
authors.lastname: Arenas
authors.affiliation: Division of Cardiology, Department of Medicine, Karolinska Institutet, 17177 Stockholm, Sweden.
authors.forename: Christer
authors.initials: C
authors.lastname: Sylvén
authors.affiliation: Division of Matrix Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 17177 Stockholm, Sweden; Duke-NUS Graduate Medical School, Durham, NC 27710, USA.
authors.forename: Karl
authors.initials: K
authors.lastname: Tryggvason
authors.affiliation: Division of Cardiothoracic Surgery and Anesthesiology, Department of Molecular Medicine and Surgery, Karolinska Institutet, 17177 Stockholm, Sweden.
authors.forename: Matthias
authors.initials: M
authors.lastname: Corbascio
authors.affiliation: Division of Cardiothoracic Surgery and Anesthesiology, Department of Molecular Medicine and Surgery, Karolinska Institutet, 17177 Stockholm, Sweden.
authors.forename: Oscar E
authors.initials: OE
authors.lastname: Simonson
authors.affiliation: Division of Cardiothoracic Surgery and Anesthesiology, Department of Molecular Medicine and Surgery, Karolinska Institutet, 17177 Stockholm, Sweden; Cell Therapy Institute, Nova Southeastern University, Fort Lauderdale, FL 33314, USA.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Österholm
authors.affiliation: Division of Cardiothoracic Surgery and Anesthesiology, Department of Molecular Medicine and Surgery, Karolinska Institutet, 17177 Stockholm, Sweden; Cell Therapy Institute, Nova Southeastern University, Fort Lauderdale, FL 33314, USA. Electronic address: karl-henrik.grinnemo@karolinska.se.
authors.forename: Karl-Henrik
authors.initials: KH
authors.lastname: Grinnemo
mesh_terms: D002319:Cardiovascular System / Q000166:cytology; D002454:Cell Differentiation / Q000235:genetics; D049109:Cell Proliferation / Q000235:genetics*; D002478:Cells, Cultured; D005318:Fetal Heart / Q000166:cytology; D020869:Gene Expression Profiling / Q000379:methods; D000072576:Homeobox Protein Nkx-2.5 / Q000235:genetics / Q000378:metabolism; D006801:Humans; D060850:LIM-Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D007797:Laminin / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism; D060449:Wnt Signaling Pathway / Q000235:genetics*; D051176:beta Catenin / Q000235:genetics* / Q000378:metabolism
chemical_list: D000072576:Homeobox Protein Nkx-2.5; D060850:LIM-Homeodomain Proteins; D007797:Laminin; C095552:NKX2-5 protein, human; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D014157:Transcription Factors; D051176:beta Catenin; C063843:insulin gene enhancer binding protein Isl-1; D040301:Vascular Endothelial Growth Factor Receptor-2