Accutase solution

Transduced TPC-1 and 8505C cells were plated at a density of 15000 cells/cm² in 12-well dishes. The next day, cell cultures were refed with fresh medium. Measurement of cell culture density was performed over 7 days after plating. Cells were detached by incubation with accutase solution (PAA Laboratories). After vigorous pipetting, the amounts of cells in suspension were determined using an electronic cell counter (model Z1 Coulter counter, Beckman Coulter).

Two cell lines from human thyroid cancers were studied: TPC-1 derived from papillary carcinoma and 8505C derived from anaplastic carcinoma (kindly gifted by Professor C. Maenhaut, IRIBHM ULB, Brussels, Belgium). The HeLa cell line derived from a cervical carcinoma (kindly gifted by Dr J. Martial, ULg, Liège, Belgium) was used as the positive control of galectin-1 expression. The TPC-1 and 8505C cell lines were grown in RPMI-1640 (Lonza, Verviers, Belgium) supplemented with 10% fetal bovine serum (FBS, Lonza, Verviers, Belgium) and 1% penicillin/streptomycin (PAA Laboratories, Pasching, Austria). HeLa cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% FBS and 1% penicillin/streptomycin. Cells were incubated at 37°C in a humidified 95% air/5% CO₂ atmosphere. The culture medium was changed once every two days. For routine maintenance and experimental studies, cells were detached by incubation with accutase solution (PAA Laboratories), resuspended and counted using an electronic cell counter (model Z1 Coulter counter, Beckman Coulter, Fullerton, CA, USA) before plating.