The presence of nifedipine in culture medium inhibits differentiation of ES cell-derived CMs.

Abstract

<p>(<b><i>A</i></b>) Protocol used to determine the effect of nifedipine application during cell differentiation. Note that for beating EBs and single cell electrophysiology and Ca2+ imaging experiments, analysis was performed at least 24 hours after washout to reflect long-term changes instead of acute drugs effects. (<b><i>B–C</i></b>) Representative experiments showing EBs with GFP expressing areas cultured under control conditions (<i>B</i>) and after nifedipine-treatment (10 µM) (<i>C</i>) of ES cells. (<b><i>D–F</i></b>) Representative EBs showing EGFP+ CMs differentiated of ES cells under control conditions (<b><i>D</i></b>), after nifedipine- (<b><i>E</i></b>) and BayK8644- (<b><i>F</i></b>) treatment. (<b>G</b>) Representative FACS analysis of ES cell-derived CMs generated under control, nifedipine- and BayK8644-treated conditions. (Scale bars 50 µm (B, C) and 20 µm (D–F).</p