Snu1544

The human colorectal cancer cell lines HCT116, DLD-1, LS174T, LS180, HCT-15, SW948, T84, Mip101, were obtained from American Type Culture Collection and SNU1544 was obtained from the Korean Cell Line Bank, and identities were confirmed by DNA profiling at the University of Colorado Cancer Center DNA Sequencing and Analysis Core. Cells were cultured in Roswell Park Memorial Institute (RPMI) media, supplemented with 10% FBS (Invitrogen), 1% nonessential amino acids (Cellgro Mediatech), 1% penicillin/streptomycin, and 0.1% puromycin. Cells were maintained in an incubator at 37° in 5% CO₂.

The human colon carcinoma CEA-positive, onalespib-sensitive cell line HT55 was obtained from the European Collection of Authenticated Cell Culture (ECACC [43 (link)]), and the colon adenocarcinoma CEA-positive, onalespib-sensitive cell line SNU1544 was obtained from Korean Cell Line Bank (KCLB [44 (link),45 (link)]). CEA expression level and onalespib sensitivity in both cell lines have been previously assessed [27 (link),46 (link)]. The HT55 cells were cultured in Minimum Essential Medium (MEM) (Biowest, Riverside, MO, USA) supplemented with 20% (v/v) fetal bovine serum (FBS; Sigma Aldrich, St. Louis, MO, USA). The SNU1544 cell line was cultured in RPMI [(Biowest, MO, USA, containing 25 nM N-2-Hydroxyethylpiperazine-N’-2-Ethanesulfonic Acid (HEPES)] supplemented with 10% (v/v) heat-inactivated FBS (Sigma Aldrich, MO, USA). Heat inactivation was done at 56 °C for 30 min. All media were supplemented with antibiotics (100 IU penicillin and 100 µg/mL streptomycin, Biochrom GmbH, Berlin, Germany) and L-glutamine (Biochrom GmbH, Berlin, Germany, 2 mM). Monolayer cultures were grown in tissue culture flasks (VWR, Radnor, PA, USA) and incubated in an atmosphere containing 5% CO₂ at 37 °C. After reaching 75–85% confluency, cell passaging was performed using Trypsin-EDTA (Biochrom GmbH, Germany). All cell lines were cultured less than 3 months after purchase.