Culture Systems & Reagents for Cardiomyocyte Differentiation
Cardiomyocytes in 3 simple steps
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Generate cardiomyocytes in as few as 8 days
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:
- Harvest PSCs
- Passage PSCs in Essential 8™ Medium
- Expand PSCs in Essential 8™ Medium
- Refeed PSCs with Essential 8™ Medium
- Replace medium with Cardiomyocyte Differentiation Medium A
- Replace medium with Cardiomyocyte Differentiation Medium B
- Replace medium with Cardiomyocyte Maintenance Medium
- Refeed every other day with Cardiomyocyte Maintenance Medium
- Characterize cardiomyocytes
Figure 1. Efficiency across multiple PSC lines. Seeding density is crucial for optimal PSC cardiomyocyte differentiation. TrypLE™-dissociated PSC lines were used for setup of these studies. For two CytoTune®-derived lines, BS2 iPSC was observed to be promiscuous at higher density while differentiation of BS3 iPSC was optimal only at a specific density. For Gibco® episomal iPSC (GEPI), it was also found to be optimal at a specific density. For hESC, H9 was observed to be promiscuous at various densities. JMP® Profiler tool identified optimal seeding density for efficient differentiation of different PSC lines.
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 Defferentiation Kit and labeled with Flou-4 NW Calcium Assay Kit. Cardiomyocytes were placed back into carciomyocyte mainteance media and imaged on EVOS® FL Auto at 4x magnification using LiteCam HD software capturing 30 frames per second.
- Efficiently reprogram somatic cells to pluripotent stem cells (PSCs) with the CytoTune®-iPS 2.0 Reprogramming Kit.
- Culture PSCs feeder-free with Essential 8™ Medium.
- Generate beating cardiomyocytes in as few as 8 days with the PSC Cardiomyocyte Differentiation Kit.
- Effectively visualize cardiomyocyte markers with the tools you need in one box.
- Alkaline Phosphatase Live Stain
- Pluripotent Stem Cell 4-Marker Immunocytochemistry Kit
- Pluripotent Stem Cell Immunocytochemistry Kit (Sox2, Tra-1-60)
- Pluripotent Stem Cell Immunocytochemistry Kit (Oct4, SSEA4)
- 3-Germ Layer Immunocytochemistry Kit
- Human Neural Stem Cell Immunocytochemistry Kit
- PSC Cardiomyocyte Differentiation Kit Prototype protocol
- Single cell dissociation of PSCs for cardiomyocyte differentiation protocol
- Reprogramming protocols
- Alkaline Phosphatase Live Stain protocol
- Video protocol: Three steps to use Alkaline Phosphatase Live Stain
- Essential 8™ protocol