Primary cell lines derived from 3 low-grade (LG) and 3 high-grade (HG) CA tissue samples included in our previous study [16 (link)] were provided by the Gillies McIndoe Research Institute Tissue Bank for this study with approval by the Central Health and Disability Ethics Committee (Ref. 15/CEN/106). Written consent was obtained from all participants. Commercial cell lines were used as positive controls for tumorsphere formation (CaCo2; cat# HTB-37, ATCC, In Vitro Technologies, Auckland, New Zealand), differentiation and α-SMA expression (3T3; cat# CRL-1658, ATCC), EpCAM expression (HepG2; cat# HB8025, ATCC) and iPSC marker expression (NTERA-2; cat# CRL-1973, ATCC).
Cells were cultured in Nunc™ EasYFlasks™ (Thermo Fisher Scientific, Waltham, MA, USA) using DMEM media with high glucose concentration and containing pyruvate (cat# 10569010, Thermo), and supplemented with 10% fetal calf serum (FCS; cat# 10091148, Thermo), 5% mTeSR Complete (cat# 85850, STEMCELL Technologies, Tullamarine, VIC, Australia), 1% penicillin/streptomycin (cat# 15140122, Thermo) and 0.2% gentamicin/amphotericin B (cat# R01510 Thermo).
Cells were passaged upon reaching 75–95% confluency using PBS (cat# 70013032, Thermo) to wash the cells and TrypLE Express Enzyme (cat# 12605093, Thermo) to detach them from the flask.
Cells were cultured in Nunc™ EasYFlasks™ (Thermo Fisher Scientific, Waltham, MA, USA) using DMEM media with high glucose concentration and containing pyruvate (cat# 10569010, Thermo), and supplemented with 10% fetal calf serum (FCS; cat# 10091148, Thermo), 5% mTeSR Complete (cat# 85850, STEMCELL Technologies, Tullamarine, VIC, Australia), 1% penicillin/streptomycin (cat# 15140122, Thermo) and 0.2% gentamicin/amphotericin B (cat# R01510 Thermo).
Cells were passaged upon reaching 75–95% confluency using PBS (cat# 70013032, Thermo) to wash the cells and TrypLE Express Enzyme (cat# 12605093, Thermo) to detach them from the flask.
Free full text: Click here
