Atcc ccl 221

DLD-1 human colorectal carcinoma cells (ATCC^® CCL-221™, Manassas, VA, USA) were obtained from the Immunology Service, Virgen de las Nieves Hospital (Granada, Spain). These types of cells were originally isolated from an adult male with a Dukes’ type C colorectal adenocarcinoma where the cancer had spread to at least one lymph node in the area close to the bowel [64 (link)].
The DLD1/Tet-On-gef, DLD1/Tet-On-apoptin, and DLD1/Tet-On-gef-apoptin cell lines, and gef and/or apoptin-expressing colorectal carcinoma cell lines controlled using a Tet-on system and induced using doxycycline (Dox) were derivates from DLD-1 cells following methodology previously described by us [16 (link)].
DLD-1 cells were grown at 37 °C, 5% CO₂ in Dulbecco’s modified Eagle medium (DMEM) (Sigma, St. Louis, MO, USA) in the presence of 10% heat-inactivated fetal bovine serum (FBS) (Sigma), 1% HEPES buffer, 40 mg/L gentamicin, 2% L-glutamine, 2.7% sodium bicarbonate and 500 mg/L ampicillin. Transfected DLD1 cell line with gef, apoptin, or both genes were cultured in the same conditions and medium described above supplemented with hygromycin B (0.4 mg/mL, Sigma) and doxycycline 0.2 mg/mL.

Human neuroblastoma SH-SY5Y (ATCC^® CRL2266™) and DLD-1 colorectal adenocarcinoma (ATCC^® CCL221™) cell lines were obtained from the American Type Culture Collection (ATCC). Human neuroblastoma SH-SY5Y cells were maintained in MEM medium and nutrient mixture F-12 Ham (1:1), supplemented with 10% foetal bovine serum, 1% sodium pyruvate and 1% glutamine. DLD-1 colorectal adenocarcinoma cell lines were maintained in RPMI media supplemented with 10% foetal bovine serum, 1% sodium pyruvate and 1% glutamine. C6-STX and C6-WT cells were obtained from the Fukuda group, Sanford-Burnham Prebys Medical Discovery Institute, La Jolla, CA, USA (for full details, see Suzuki et al.)31 (link). Briefly, C6-STX glioma cells were prepared by transfecting wild-type C6 cells (C6-WT) with the pcDNA3-STX plasmid inserted with cDNA encoded full length human ST8SiaII (also known as STX)31 (link). Both cell lines were maintained in minimum essential medium (MEM Alpha Eagle) with ultraglutamine I, deoxyribonucleoside and ribonucleosides supplemented with 10% foetal bovine serum. All cell lines were maintained in a humid atmosphere of 5% CO₂ and 95% air at 37 °C.
For studies carried out under conditions of low oxygen tension (“hypoxic conditions”), all media used was incubated for at least 48 hours under hypoxic conditions (0.1% oxygen) before being added to the cells.