4.7 Article

Scalable Electrophysiological Investigation of iPS Cell-Derived Cardiomyocytes Obtained by a Lentiviral Purification Strategy

Journal

JOURNAL OF CLINICAL MEDICINE
Volume 4, Issue 1, Pages 102-123

Publisher

MDPI
DOI: 10.3390/jcm4010102

Keywords

induced pluripotent stem cells; cardiomyocytes; purification; long QT syndrome; planar patch clamp; microelectrode array

Funding

  1. German Research Foundation [SA 1785/5-1]
  2. StemCellFactory project
  3. European Union (European Regional Development Fund-Investing in your future)
  4. German federal state, North Rhine-Westphalia (NRW)

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Disease-specific induced pluripotent stem (iPS) cells can be generated from patients and differentiated into functional cardiomyocytes for characterization of the disease and for drug screening. In order to obtain pure cardiomyocytes for automated electrophysiological investigation, we here report a novel non-clonal purification strategy by using lentiviral gene transfer of a puromycin resistance gene under the control of a cardiac-specific promoter. We have applied this method to our previous reported wild-type and long QT syndrome 3 (LQTS 3)-specific mouse iPS cells and obtained a pure cardiomyocyte population. These cells were investigated by action potential analysis with manual and automatic planar patch clamp technologies, as well as by recording extracellular field potentials using a microelectrode array system. Action potentials and field potentials showed the characteristic prolongation at low heart rates in LQTS 3-specific, but not in wild-type iPS cell-derived cardiomyocytes. Hence, LQTS 3-specific cardiomyocytes can be purified from iPS cells with a lentiviral strategy, maintain the hallmarks of the LQTS 3 disease and can be used for automated electrophysiological characterization and drug screening.

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