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

Abstract

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.

Comparing human iPSC-cardiomyocytes versus HEK293T cells unveils disease-causing effects of Brugada mutation A735V of NaV1.5 sodium channels

“…1e). IF staining of cardiac aggregate-derived dissociated CMs cultured for 29 days on glass coverslips revealed (i) prominent expression of Na V 1.5 comparable across WT and mutant hiPSC-CMs with speckled channel distribution patterns confirming previously published confocal images of Na V 1.5 in neonatal 23 and iPSC-derived cardiomyocytes 24,25 (ii) prominent ß-MyHC isoform expression (encoded by MYH7 ; α- to ß-MyHC isotype switch being an accepted maturation mark in the native heart and in hPSC-derived CMs as well 19 ) and (iii) induction of organized sarcomere structures and abundant CMs’ bi-nucleation (Fig. 1f).…”

Frequency-dependent drug screening using optogenetic stimulation of human iPSC-derived cardiomyocytes

“…Furthermore, the large stimulation artefacts at the beginning of the FP signals makes the analysis of the initial rapid FP component impossible. This is why most studies of cardiomyocytes on MEA are done during spontaneous beating 1 , 6 , 10 , 20 , 22 – 25 , 29 , 30 . However drug effects on cardiac excitability and action potentials can depend on the heart rate because of use-dependent properties of cardiac ion channels and the rate-dependency of drugs 31 – 33 .…”

Convergences of Life Sciences and Engineering in Understanding and Treating Heart Failure

“…Small animal models are commonly used in cardiovascular research because of their many advantages over large animal models including a short life span that allows the investigators to follow the natural history of the disease at an accelerated pace. Also, the use of genetically modified animals allows for rigorous mechanistic studies that can be further validated in larger animal models 26 . These in vivo studies are not only mandatory to test the safety and efficacy of stem cells but also to assess how bioengineering can optimize outcomes through improvement of cell survival and engraftment and/or long-lasting release of cell-secreted factors (e.g.…”