Healthy human iPSC line characterization

Human iPSC line (Ctrl-2), derived from a healthy mid-age Caucasian female donor peripheral blood mononuclear cells (PBMCs), established and characterized earlier [32 (link),33 (link)], was used in this study. Cells were cultured on BD Matrigel™ matrix (BD Biosciences, Franklin Lakes, NJ, USA) with mTeSR™1 medium (Stem Cell Technologies, Vancouver, Canada), using Gentle Cell Dissociation Reagent for passages, according to the manufacturer’s instruction. Representative hiPSC colony morphology, pluripotency staining for POU5F1, NANOG, SSEA4, and TRA1-60 and karyotype analysis are presented in Figure S1. For mycoplasma screening, the Venor^®GeM-Advance (Minerva Biolabs) Mycoplasma Detection Kit was used according to the manufacturer’s protocol in every fifth passage during maintenance and before freezing. Cells were cultured at 37 °C in a humidified atmosphere containing 5% CO₂. In the current study cultures from passage 16 and 17 were used for differentiation.