Cardiomyocytes in 3 simple steps
In a recent collaboration, scientists at Stem Cell Theranostics used Thermo Fisher Scientific workflow reagents to develop stem cell-derived cardiomyocytes from patients with inherited hypertrophic cardiomyopathy (HCM).
View the white paper describing their protocols and results
For more difficult to differentiate iPSC lines, our supplemental enrichment protocol will enable 10–30% greater efficiency. Cells can be characterized after differentiation using the Invitrogen Human Cardiomyocyte Immunocytochemistry Kit.
View the enrichment protocol
A simple protocol is used to initiate PSC induction to cardiomyocytes in as few as 8 days.
The protocol can be viewed on the left and includes:
Figure 1. Efficiency across multiple PSC lines. TrypLE-dissociated PSC lines were seeded at specific density onto Geltrex-coated surface and cultured in Essential 8 media. After three days expansion, PSC lines at optimal confluency were induced using the PSC Cardiomyocyte Differentiation Kit according to protocol and cultured for two weeks. Cells were harvested and analyzed for TNNT2+ cells by flow cytometry. Results showed high cardiomyocyte differentiation efficiency among all lines when it reaches optimal confluency at time of induction.
Figure 2. iPSCs were differentiated for 14 days using the PSC Cardiomyocyte Differentiation Kit. The cells were stained using the Human Cardiomyocyte Immunocytochemistry Kit for the following markers: NKX2.5 (red) for early cardiac mesoderm and TNNT2/cTNT (green) for cardiomyocytes along with DAPI nuclear DNA counterstaining (blue).
Figure 3. Cardioactive compounds can modulate PSC-derived cardiomyocyte contraction. Here we show the response of spontaneous beat rate on cardiomyocytes differentiated using PSC Cardiomyocyte Differentiation Kit measured using Multi Electrode Array. (A) Cardiomyocyte waveform under baseline conditions. (B) Cardiomyocyte waveform under verapamil treatment conditions. (C) Effect of verapamil, an L-type Ca2+ channel blocker, on the spontaneous beat rate of H9-derived cardiomyocytes; at the highest dose level, cardiomyocyte contraction ceased. (D) Effect of isoproterenol, a beta-adrenergic receptor agonist, on the spontaneous beat rate of H9-derived cardiomyocytes.
Figure 4. H9 cells differentiated into cardiomyocytes using PSC Cardiomyocyte Differentiation Kit and labeled with Fluo-4 NW Calcium Assay Kit. Cardiomyocytes were placed back into cardiomyocyte maintenance media and imaged on EVOS FL Auto at 4x magnification using liteCam HD software capturing 30 frames per second.