Venor gem qonestep kit

hiPSCs were cultured in mTeSR™1 medium (Stemcell Technologies, Vancouver, BC, Canada) on Matrigel-coated (Corning™ hESC-qualified) 6-well plates. hiPSCs were cultured in hypoxic conditions at 5% O₂ as this was shown to improve hiPSC quality [50 (link),51 (link)]. Cells were passaged routinely at a ratio of 1:10 every 3 days using 0.5 mM PBS/EDTA. Mycoplasma was tested using the Venor^® GeM qOneStep kit (Minerva Biolabs, Berlin, Germany).

The human breast cancer cell lines MCF-7, SK-BR-3 and MBA-MB-231 (triple-negative), LNCap (human prostate carcinoma lymph node metastasis), HeLa (cervix carcinoma), HepG2 (hepatocellular carcinoma), and HCT116 (human colon carcinoma) were purchased commercially from DSMZ (Braunschweig, Germany). If not otherwise stated, all cell culture media and supplements were obtained from Life Technologies (Darmstadt, Germany).
Cells were cultured as described previously [36 (link)]. In detail, MCF-7 in RPMI medium containing 10% fetal bovine serum (FBS), 1 mM sodium pyruvate, and 0.1 M nonessential amino acids (NEAA); HeLa and HepG2 in RPMI medium containing 10% FBS and 2 mM Glutamax; MDA-MB-231 in Leibovitz’s L15 with 10% FBS; HCT116 and SK-BR-3 in McCoy’s 5A medium containing 10% FBS and 2 mM Glutamax; and LNCap in RPMI medium containing 10% FBS, 1 mM sodium pyruvate, 0.1 M NEAA, 2 mM Glutamax, and 0.01 M HEPES. All cell culture media were used without antibiotics. MCF-7, HCT116, SK-BR-3, LNCap, HeLa, and HepG2 were maintained at 37 °C in a humidified incubator with a 5% CO₂ atmosphere and MDA-MB-231 cells were handled without gaseous exchange. The absence of mycoplasma in the cell culture was regularly tested using the Venor GeM qOneStep-Kit (Minerva-biolabs, Berlin, Germany).