Maintenance and Differentiation of iPSC-CMs

iPSC-CM were commercially obtained directly from Axiogenesis [note this company has recently merged with Pluriomics to form NCardia] (#Ax-B-HC02-1M) and Cellular Dynamics International (CDI; #CMC-100-012-000.5). Upon receipt, cells were transferred to liquid nitrogen for long term storage. Thereafter, when required for use, cells were retrieved and plated according to manufacturer’s instructions into wells of a 96-well plate coated with fibronectin from bovine plasma (Sigma) at a final concentration of ~2 μg/cm². Axiogenesis and CDI iPSC-CM were plated as recommended at a density of 35,000 or 50,000 viable cells per well respectively, unless stated otherwise. Cells were maintained in a sterile humidified incubator (37°C, 5% CO₂) in the appropriate maintenance medium provided by the manufacturers, and was replaced 4–24 hours post plating, and every 24–48 hours thereafter.
3T3-L1 fibroblasts were obtained directly from ATCC (#CL-173) and grown and differentiated as outlined previously [25 (link)].