Growth factor reduced matrigel coated

Experiments were performed using the human induced pluripotent stem cell (hiPSC) line UMGi014-C clone 14 (isWT1.14) provided by LC (Stem Cell Unit, University Medical Center Göttingen). The iPSC generation and characterization for pluripotency and genomic stability was described previously (18 (link)). The line was reprogrammed from somatic cells of a healthy 35-year-old Caucasian male individual. Human iPSC cultures were maintained in Stem MACS iPS Brew XF Medium (Miltenyi Biotec; #130-104-368) in 17 μg/cm² Growth Factor Reduced Matrigel-coated (Corning; #354230) 6-well dishes in a humidified normoxic incubator (37°C, 5% CO₂). Cultures were routinely passaged in colonies at a ratio of 1:16 every 4–5 days after dissociation using Versene Solution [0.48 mM ethylenediamine tetraacetic acid (EDTA)] (Thermo Fisher; #15040033). Cells were plated in Stem MACS iPS Brew XF Medium and 1 μl/ml of 2 mM Thiazovivin (Millipore; #420220) in DMSO (final concentration 2 μM) for the first 24 h. The culture medium was changed daily with 2 ml per well Stem MACS iPS Brew XF Medium.

The hiPSC cell line (CS83iCTR-33n1) was provided by the Cedars-Sinai Medical Center’s David and Janet Polak Foundation Stem Cell Core Laboratory. These cells have been generated through episomal reprogramming of fibroblasts of a 31-year-old healthy female. The cell line has been fully characterized and no karyotype abnormalities have been found. hiPSCs were cultured on growth factor-reduced matrigel-coated (Corning) cell culture plates in mTeSR1 medium (Stem Cell Technologies, Vancouver, Canada) and were passaged using gentle cell dissociation reagent (Stem Cell Technologies).