Cell Culture and Maintenance of Diverse Cell Lines

Calu-3 cells derived from the pleural effusion of a 25 years old patient with a lung adenocarcinoma [48 (link)] and VeroE6 cells, immortalized African green kidney epithelial cells [49 ], (both provided by Prof. Robert Rieben, Department of Biomedical Research (DBMR), University of Bern) were grown in Dulbecco’s modified Eagle Medium (DMEM)–GlutaMAX™, 10% (v/v) heat-inactivated fetal bovine serum (FBS), 100 mg/mL streptomycin, 100 IU/mL penicillin, 1% (w/v) non-essential amino acids and 15 mM HEPES (all Gibco). HIBCPP (a cell line obtained from a choroid plexus papilloma [47 (link)]) were maintained in DMEM/F12, 10% v/v heat-inactivated FBS (Gibco), 5 µg/mL human insulin (Sigma Aldrich), 4mM L-Glutamine (Gibco) and 100 mg/mL streptomycin, 100 IU/mL penicillin (HIBCPP-10 medium). Human induced pluripotent stem cells (hiPSCs) were generated from erythroblasts reprogrammed through nucleofection with plasmids encoding for OCT4, shRNAp53, SOX2, KLF4, L-Myc, and Lin28 [50 (link)]. In this study 2 iPSC clones per donor were used: LNISi001-A/B, LNISi002-A/B, LNISi003-A/B, LNISi004-A/B, with the following age/sex: 30/female, 50/male, 49/female, 33/female. HiPSCs were maintained on growth factor reduced Matrigel^® (Corning) or growth factor reduced and stem cell certified Cultrex™ (R&D) coated plates in mTeSR^TM1 medium (STEMCELL Technologies) [42 (link), 51 (link)].