Stemdiff cm differentiation kit

All studies were performed under the regulation of the SickKids Research Ethics Board and Canadian Institutes of Health Research Stem Cell Oversight Committee. Human iPS cells derived from peripheral blood lymphocytes of a healthy adult male donor (PGPC-17),¹⁰⁶ (link) were differentiated into cardiomyocytes (CMs) using the STEMdiff CM Differentiation Kit (STEMCELL Technologies). hTERT-immortalized female adipose-derived primary human mesenchymal stem cells (MSCs, SCRC4000, ATCC) were maintained in basal MSC media (PCS-500-030, ATCC), supplemented with 2 % FBS (ThermoFisher), 5 ng/ml recombinant human FGF basic (R&D Systems 233-FB-010), 5 ng/ml recombinant human FGF acidic (R&D Systems 232-FA-025), 5 ng/ml recombinant human EGF (R&D Systems 236-EG-200), 2.4 mM L-Alanyl-L-Glutamine (ThermoFisher), and 0.2 mg/ml Geneticin (G418, ThermoFisher). MSCs were differentiated into VSMCs over the course of 19 days by supplementing the above described basal MSC media with 1 ng/ml TGF-β1 (R&D Systems 240-B-002) as previously described.¹²⁴ (link)^,125 (link) HEK-293 cells (ATCC) were cultured in EMEM (Gibco), supplemented with 10 % FBS (Canadian origin, ThermoFisher) and 1 % Penicillin/Streptomycin (ThermoFisher). All cells were maintained at sub-confluent conditions, maintained at 37 C with 5 % CO₂ and were passaged every 3-4 days.