title: Modeling Cardiovascular Diseases with Patient-Specific Human Pluripotent Stem Cell-Derived Cardiomyocytes.
abstract: The generation of cardiomyocytes from human induced pluripotent stem cells (hiPSCs) provides a source of cells that accurately recapitulate the human cardiac pathophysiology. The application of these cells allows for modeling of cardiovascular diseases, providing a novel understanding of human disease mechanisms and assessment of therapies. Here, we describe a stepwise protocol developed in our laboratory for the generation of hiPSCs from patients with a specific disease phenotype, long-term hiPSC culture and cryopreservation, differentiation of hiPSCs to cardiomyocytes, and assessment of disease phenotypes. Our protocol combines a number of innovative tools that include a codon-optimized mini intronic plasmid (CoMiP), chemically defined culture conditions to achieve high efficiencies of reprogramming and differentiation, and calcium imaging for assessment of cardiomyocyte phenotypes. Thus, this protocol provides a complete guide to use a patient cohort on a testable cardiomyocyte platform for pharmacological drug assessment.
authors.affiliation: Department of Medicine (Division of Cardiology), Stanford Cardiovascular Institute, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA. burridge@stanford.edu.
authors.forename: Paul W
authors.initials: PW
authors.lastname: Burridge
authors.affiliation: Max Delbrück Center, Berlin Institute of Health, Berlin, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Diecke
authors.affiliation: Department of Medicine (Division of Cardiology), Stanford Cardiovascular Institute, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA.
authors.forename: Elena
authors.initials: E
authors.lastname: Matsa
authors.affiliation: Department of Medicine (Division of Cardiology), Stanford Cardiovascular Institute, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA.
authors.forename: Arun
authors.initials: A
authors.lastname: Sharma
authors.affiliation: Department of Medicine (Division of Cardiology), Stanford Cardiovascular Institute, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA.
authors.forename: Haodi
authors.initials: H
authors.lastname: Wu
authors.affiliation: Department of Medicine (Division of Cardiology), Stanford Cardiovascular Institute, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, 265 Campus Drive, Room G1120B, Stanford, CA, 94305-5454, USA. joewu@stanford.edu.
authors.forename: Joseph C
authors.initials: JC
authors.lastname: Wu
mesh_terms: D000954:Antigens, Surface / Q000235:genetics / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002311:Cardiomyopathy, Dilated / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D002454:Cell Differentiation; D065150:Cellular Reprogramming; D015925:Cryopreservation; D020405:Dermis / Q000166:cytology / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D008954:Models, Biological; D057054:Molecular Imaging; D009154:Mutation; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D061251:Primary Cell Culture; D011509:Proteoglycans / Q000235:genetics / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D012692:Serial Passage; D020107:Troponin T / Q000235:genetics / Q000378:metabolism
chemical_list: D000954:Antigens, Surface; D015415:Biomarkers; D036341:Intercellular Signaling Peptides and Proteins; D011509:Proteoglycans; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; C545568:TNNT2 protein, human; C107064:TRA-1-60 antigen, human; D020107:Troponin T; D002118:Calcium
keywords: Calcium imaging; Cardiomyocytes; Disease modeling; Human induced pluripotent stem cells