Evaluating Cardiac Effects of GLP-26 in Human Cardiomyocytes

Freshly thawed cryopreserved human cardiomyocytes were seeded in 24-well collagen-coated plates with iCell plating medium at a density of 240,000 (plating efficiency 53%) and incubated in a cell culture incubator at 37 °C, 7% CO₂. After 48 h, non-adhered cardiomyocytes and debris were removed by rinsing twice with iCell maintenance medium and the attached cells were incubated for an additional eight days in maintenance medium, with fresh medium replacement every other day. Ten days post-plating when all cells demonstrated regular synchronous beating, cardiomyocytes were then exposed to 2 and 10 µM (similar to the maximal total (free + bound) plasma concentration) of GLP-26 prepared in DMSO. The same volume of DMSO was used as negative control, and 50 µM of BMS-986094 was used as positive control [21 (link)]. Cultures were imaged using a PerkinElmer spinning disc confocal microscope (PerkinElmer, Waltham, MA, USA) and 30 s videos were recorded at 14 fps using a Hamamatsu Flash 4.0 sCMOS camera and Velocity Software at 2, 4, 8, 12, 24, and 48 h after drug exposure. Automated video-based analysis of contractility by a computational motion tracking software was applied to identify potential cardiac effects of GLP-26 [22 (link)].