Generation of highly purified human cardiomyocytes from peripheral blood mononuclear cell-derived induced pluripotent stem cells

• Published in: PloS one 2015-05, Vol.10 (5), p.e0126596-e0126596
• Main Authors: Fuerstenau-Sharp, Maya, Zimmermann, Martina E, Stark, Klaus, Jentsch, Nico, Klingenstein, Melanie, Drzymalski, Marzena, Wagner, Stefan, Maier, Lars S, Hehr, Ute, Baessler, Andrea, Fischer, Marcus, Hengstenberg, Christian
• Format: Article
• Language: English
• Subjects: Action Potentials; Antibodies; B cells; Blood; Calcium Signaling; Calcium-binding protein; Cardiomyocytes; Cell culture; Cell Differentiation; Cells, Cultured; Cellular Reprogramming; Differentiation; Drug discovery; Drug testing; Epidemiology; Equipment costs; Fibroblasts; Flow Cytometry; Gap Junctions; Glucose; Growth factors; Heart; Heart cells; Heart diseases; Hospitals; Humans; Immunomagnetic Separation; Induced Pluripotent Stem Cells - physiology; Internal medicine; Karyotype; Lactates; Lactic acid; Long QT syndrome; Lymphocytes; Lymphocytes T; Myocytes, Cardiac - physiology; Myosin; Patch-Clamp Techniques; Peripheral blood; Pluripotency; Preventive medicine; Regenerative medicine; Retroviridae - genetics; Stem cells; T cells; T-Lymphocytes - physiology; Troponin; Troponin T; Ventricle; α-Actinin
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0126596

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.