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(A) High cardiac purity by cTNT flow cytometry, and (B) High viability as indicated by CyQUANT Direct Cell Proliferation Assay (normalized to maximal response per experiment and reported as % iCMM) at end of recommended workflow: D14 cultured in iCMM or iCSFM; D11 cultured in iCTAM. (Purity n=3-6; Viability n= 64-90). (C) Using the xCELLigence CardioRTCA, iCell Cardiomyocytes2 cultured in iCSFM or iCTAM maintained comparable cell index (measure of cell attachment and health) and beat rate (not shown) to those cultured in iCMM (D7 n=24-48; D11 n=9-18). (D) Using the Maestro Pro MEA platform, iCell Cardiomyocytes2 cultured in iCSFM or iCTAM maintained comparable percent total active electrodes (not shown), beat rate, low beat rate irregularity, and field potential duration (FPD corrected to beat rate) to those cultured in iCMM. We recommend iCTAM for up to 4 days as we observed a reduction in beat rate with cells cultured in iCTAM for 7d. (n=24-48)
Representative traces for (A) Impedance measurements (xCELLigence), (B) Field potential (with T-wave marked) and (C) Action potential (LEAP) traces from Day 7, 4h after media change. Note: In (B) and (C) the cells had been treated with 0.1% DMSO vehicle control.
(A) Comparison of mean beats are shown of iCell Cardiomyocytes2 cultured in iCell Cardiomyocytes Serum-free Medium (iCSFM) and serum containing iCell Cardiomyocytes Maintenance Medium (iCMM) 24 hours after medium switch. (B) A bar graph showing contractile parameters (amplitude, beat rate, beat duration, area under curve, upstroke and downstroke slopes) of iCell Cardiomyocytes² cultured in iCell Cardiomyocytes Serum-free Medium after 24 hours. Data is normalized to iCell Cardiomyocytes² cultured in iCell Cardiomyocytes Maintenance Medium. (Data kindly provided by Innovitro and Nanion Technologies)
Representative LEAP traces recorded from iCell Cardiomyocytes2 treated with DMSO (control) or Drug (Nifedipine (50nM) top panels, E4031 (30nM), bottom panel) when cultured in iCMM (left), iCSFM (middle) or iCTAM (right). iCell Cardiomyocytes2 were thawed and plated according to User's Guide and cultured until DIV7 in iCMM. On DIV7 some of the wells were washed and fed with iCSFM (for 4 hours and then dosed) or iCTAM (cultured until DIV11 and then dosed).