Fuerstenau-Sharp, Maya and Zimmermann, Martina E. and Stark, Klaus and Jentsch, Nico and Klingenstein, Melanie and Drzymalski, Marzena and Wagner, Stefan and Maier, Lars S. and Hehr, Ute and Baessler, Andrea and Fischer, Marcus and Hengstenberg, Christian (2015) Generation of Highly Purified Human Cardiomyocytes from Peripheral Blood Mononuclear Cell-Derived Induced Pluripotent Stem Cells. PLOS ONE, 10 (5): UNSP e0126. ISSN 1932-6203,
Full text not available from this repository. (Request a copy)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, alpha-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 cardio-myocyte 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.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | LONG-QT SYNDROME; ACTION-POTENTIALS; PROGENITOR CELLS; T-CELLS; MOUSE; CONTRACTION; DIFFERENTIATION; PURIFICATION; CURRENTS; MYOCYTES; |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Humangenetik Medicine > Lehrstuhl für Innere Medizin II Medicine > Institut für Epidemiologie und Präventivmedizin |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 15 Jul 2019 09:07 |
| Last Modified: | 15 Jul 2019 09:07 |
| URI: | https://pred.uni-regensburg.de/id/eprint/5504 |
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