Discover Your Next iPSC-derived Cardiomyocytes.NEXEL Co., Ltd. strives to provide high quality human cardiomyocytes derived from human induced pluripotent stem (iPS) cells using optimized proprietary protocols. The Cardiosight®-S is a highly pure and electrophysiologically active population of cells, suitable for all types of experiments in the field of cardiomyocytes. Our priority is to ensure researchers get a reliable product and accelerate the diverse projects they are working on.
Our human iPSC-derived Cardiomyocytes have been validated on multiple platforms and our quality control process has been optimized to test the exact settings in which customers use our product. Through and comprehensive testing ensures that the cells have been tested both on the Nanion Cardioexcyte96 and the Axion Maestro MEA, making them the best choice for your MEA assay. Our Certificate of Analysis (CoA) contains the exact results we obtained and our technical support team will work with you to provide the same experience in your lab.
The Cardiosight®-S recovers quickly upon thawing to start spontaneous beating the 2 days after and stabilizing over time (45~60 bpm on DIV7). The Cardiosight®-S cardiomyocytes readily form a single synchronous monolayer which enhances both reproducibility and accuracy of assays. The above images were taken from a 12-well culture plate 72 hours, 1 week and 2 weeks post-thawing (Magnification 100x). Please note that the Cardiosight®-S was seeded following the Cardiosight®-S User Guide but at a plating density of 150,000 cells/㎠ to demonstrate correct layer formation even at lower densities. Specific applications such as MEA assays are optimized to different densities and will not display the same morphology as here.
Immunostaining
100x
DIV 7
MLC2V
MLC2A
Merged
Connexin43
cTnT
Merged
The Cardiosight®-S expresses typical ventricular cardiomyocyte markers and structural characteristics which are indicative of appropriate contractile functionality of these cells. In particular, the Cardiosight®-S demonstrates high expression of Connexin 43, a gap junction protein, which suggests high interconnectivity and thus synchronicity of the myocytes a must for good MEA assay results. 30,000 cells were seeded on Matrigel on a 9 mm coverslip and cultured for 7 days before fixation. Immunocytochemistry was performed by incubating primary antibodies at the concentrations recommended by the respective suppliers at RT for 1 hour.
The Cardiosight®-S proved a good fit with the Nanion Patchliner for recording patch clamp measurements of the sponataneous beating. The cells demonstrated a high success rate and low variability in the sodium currents (INa). It was possible to measure a dose-dependent block of sodium currents by TTX (A) and effects specific to INa, Late by ATX-II (enhancer) and Ranolazine (blocker) were also easily observable. Finally, calcium channel activation by BayK and inhibition by Nifedipine could also be detected in changes of the APD90. These results demonstrate the Cardiosight®-S expresses the correct ion channels and can readily be used in pharmacology applications. Read our poster here and find out more about the Nanion Patchliner Dynamite8here.
The Cardiosight®-S demonstrated great results also in dynamic clamp experiments, again on the Nanion Patchliner with the Dynamite8 add-on. In A, it was possible to record adult-ventricular-like action potentials with the the Cardiosight®-S at physiological temperature. In B & C, the effects of calcium channel activation (BayK) and inhibition (Nifedipine) could be assessed also in the presence of dynamic clamping. Read our poster here and find out more about the Nanion Patchliner here.
The Cardiosight®-S demonstrated low inter-well variability when tested on the Nanion CardioExcyte 96. High reproducibility of Impedance and EFP signals facilitated compound testing experiments (Nifedipine & Dofetilide). Concentrations were increased by consecutive treatments of 10% media changes and matched the expected results, validating the cardiomyocyte properties. Read our poster here and find out more about the Nanion CardioExcyte 96 here.
On Nanion’s all-new FLEXcyte 96 which uses soft subtrates to mature the cardiomyocytes, the Cardiosight®-S demonstrated high responsiveness to Isoproterenol and were able to detect positive inotropic effects after just 7 days of culture. Find out more about the Nanion FLEXcyte here.
Electrophysiology – Axion Maestro
100x
DIV 1
DIV 3
DIV 5
DIV 7
DIV 11
DIV 14
The above images were taken from a 48well Axion MEA plate 1st, 3rd, 5th, 7th, 11th, 14th day post-thawing (Magnification 100x). Please note that the Cardiosight®-S was seeded following the Cardiosight®-S Maestro Application Protocol.
Nifedipine (CaV blocker)
E4031 (hERG blocker)
Dofetilide (hERG blocker)
Mexiletine (Sodium channel blocker)
The Cardiosight®-S demonstrated low variability, good synchronicity and good compound reactivity(Nifedipine, E4031, Dofetilide, Mexiletine) on the Axion Maestro. NEXEL’s QC process makes sure every batch of cells behaves similarly and the above data is available for each vial upon request. We also make sure these results are reproducible independently of which platform you choose to do the MEA assay on.