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

PLoS One. 2015 May 13;10(5):e0126596. doi: 10.1371/journal.pone.0126596. eCollection 2015.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Action Potentials
  • Calcium Signaling
  • Cell Differentiation
  • Cells, Cultured
  • Cellular Reprogramming
  • Flow Cytometry
  • Gap Junctions
  • Humans
  • Immunomagnetic Separation
  • Induced Pluripotent Stem Cells / physiology*
  • Karyotype
  • Myocytes, Cardiac / physiology*
  • Patch-Clamp Techniques
  • Retroviridae / genetics
  • T-Lymphocytes / physiology*

Grants and funding

This work was supported by the National Genome Network (NGFNplus Atherogenomics, 01GS0831) (CH, MFS), and the European Union 6th Framework Program Cardiogenics (LSH-2006-037593) (CH). SW and LSM are funded by Deutsche Forschungsgemeinschaft (DFG) through an International Research Training Group GRK 1816. LSM is funded by the Fondation Leducq Transatlantic Network on “Redox and Nitrosative Regulation of Cardiac Remodeling”.