title: Patient-Specific Induced Pluripotent Stem Cell Models: Generation and Characterization of Cardiac Cells.
abstract: The generation of human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes has been of utmost interest for the study of cardiac development, cardiac disease modeling, and evaluation of cardiotoxic effects of novel candidate drugs. Several protocols have been developed to guide human stem cells toward the cardiogenic path. Pioneering work used serum to promote cardiogenesis; however, low cardiogenic throughputs, lack of chemical definition, and batch-to-batch variability of serum lots constituted a considerable impediment to the implementation of those protocols to large-scale cell biology. Further work focused on the manipulation of pathways that mouse genetics indicated to be fundamental in cardiac development to promote cardiac differentiation in stem cells. Although extremely elegant, those serum-free protocols involved the use of human recombinant cytokines that tend to be quite costly and which can also be variable between lots. The latest generation of cardiogenic protocols aimed for a more cost-effective and reproducible definition of the conditions driving cardiac differentiation, using small molecules to manipulate cardiogenic pathways overriding the need for cytokines. This chapter details methods based on currently available cardiac differentiation protocols for the generation and characterization of robust numbers of hiPSC-derived cardiomyocytes under chemically defined conditions.
authors.affiliation: Cardiology Division, Department of Medicine, University of California-San Diego, 9500 Gilman Drive, La Jolla, CA, 92093-0613C, USA.
authors.forename: Fabian
authors.initials: F
authors.lastname: Zanella
authors.affiliation: Cardiology Division, Department of Medicine, University of California-San Diego, 9500 Gilman Drive, La Jolla, CA, 92093-0613C, USA. fasheikh@ucsd.edu.
authors.forename: Farah
authors.initials: F
authors.lastname: Sheikh
mesh_terms: D000185:Actinin / Q000235:genetics / Q000378:metabolism; D000577:Amides / Q000494:pharmacology; D015415:Biomarkers / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002311:Cardiomyopathy, Dilated / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D065150:Cellular Reprogramming; D003094:Collagen / Q000737:chemistry; D004338:Drug Combinations; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D015870:Gene Expression; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D007328:Insulin / Q000494:pharmacology; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D007797:Laminin / Q000737:chemistry; D008954:Models, Biological; D057054:Molecular Imaging; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D061251:Primary Cell Culture; D011509:Proteoglycans / Q000737:chemistry; D011725:Pyridines / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D000577:Amides; D015415:Biomarkers; C473711:Chir 99021; D004338:Drug Combinations; D004791:Enzyme Inhibitors; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins; D007328:Insulin; D036341:Intercellular Signaling Peptides and Proteins; D007797:Laminin; C095552:NKX2-5 protein, human; D011509:Proteoglycans; D011725:Pyridines; D011743:Pyrimidines; D014157:Transcription Factors; D000185:Actinin; C059485:matrigel; C108830:Y 27632; D003094:Collagen; D002118:Calcium
keywords: Cardiac assays; Cardiac differentiation; Cardiomyocyte; Human induced pluripotent stem cells; Small molecule; Wnt signaling pathway