title: Generation of highly purified human cardiomyocytes from peripheral blood mononuclear cell-derived induced pluripotent stem cells.
abstract: Induced pluripotent stem (iPS) cells have an enormous potential for physiological studies. A novel protocol was developed combining the derivation of iPS from peripheral blood with an optimized directed differentiation to cardiomyocytes and a subsequent metabolic selection. The human iPS cells were retrovirally dedifferentiated from activated T cells. The subsequent optimized directed differentiation protocol yielded 30-45% cardiomyocytes at day 16 of differentiation. The derived cardiomyocytes expressed appropriate structural markers like cardiac troponin T, α-actinin and myosin light chain 2 (MLC2V). In a subsequent metabolic selection with lactate, the cardiomyocytes content could be increased to more than 90%. Loss of cardiomyocytes during metabolic selection were less than 50%, whereas alternative surface antibody-based selection procedures resulted in loss of up to 80% of cardiomyocytes. Electrophysiological characterization confirmed the typical cardiac features and the presence of ventricular, atrial and nodal-like action potentials within the derived cardiomyocyte population. Our combined and optimized protocol is highly robust and applicable for scalable cardiac differentiation. It provides a simple and cost-efficient method without expensive equipment for generating large numbers of highly purified, functional cardiomyocytes. It will further enhance the applicability of iPS cell-derived cardiomyocytes for disease modeling, drug discovery, and regenerative medicine.
authors.affiliation: Clinic for Internal Medicine II, University Hospital Regensburg, Regensburg, Germany.
authors.forename: Maya
authors.initials: M
authors.lastname: Fuerstenau-Sharp
authors.affiliation: Clinic for Internal Medicine II, University Hospital Regensburg, Regensburg, Germany.
authors.forename: Martina E
authors.initials: ME
authors.lastname: Zimmermann
authors.affiliation: Department of Genetic Epidemiology, Institute of Epidemiology and Preventive Medicine, University of Regensburg, Regensburg, Germany.
authors.forename: Klaus
authors.initials: K
authors.lastname: Stark
authors.affiliation: Clinic for Internal Medicine II, University Hospital Regensburg, Regensburg, Germany.
authors.forename: Nico
authors.initials: N
authors.lastname: Jentsch
authors.affiliation: Clinic for Internal Medicine II, University Hospital Regensburg, Regensburg, Germany.
authors.forename: Melanie
authors.initials: M
authors.lastname: Klingenstein
authors.affiliation: Clinic for Internal Medicine II, University Hospital Regensburg, Regensburg, Germany.
authors.forename: Marzena
authors.initials: M
authors.lastname: Drzymalski
authors.affiliation: Clinic for Internal Medicine II, University Hospital Regensburg, Regensburg, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Wagner
authors.affiliation: Clinic for Internal Medicine II, University Hospital Regensburg, Regensburg, Germany.
authors.forename: Lars S
authors.initials: LS
authors.lastname: Maier
authors.affiliation: Center for and Department of Human Genetics, University of Regensburg, Regensburg, Germany.
authors.forename: Ute
authors.initials: U
authors.lastname: Hehr
authors.affiliation: Clinic for Internal Medicine II, University Hospital Regensburg, Regensburg, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Baessler
authors.affiliation: Clinic for Internal Medicine II, University Hospital Regensburg, Regensburg, Germany.
authors.forename: Marcus
authors.initials: M
authors.lastname: Fischer
authors.affiliation: Clinic for Internal Medicine II, University Hospital Regensburg, Regensburg, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Hengstenberg
mesh_terms: D000200:Action Potentials; D020013:Calcium Signaling; D002454:Cell Differentiation; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D005434:Flow Cytometry; D017629:Gap Junctions; D006801:Humans; D018189:Immunomagnetic Separation; D057026:Induced Pluripotent Stem Cells / Q000502:physiology*; D059785:Karyotype; D032383:Myocytes, Cardiac / Q000502:physiology*; D018408:Patch-Clamp Techniques; D012190:Retroviridae / Q000235:genetics; D013601:T-Lymphocytes / Q000502:physiology*