Methylcellulose

Human HEMC-SS chondrosarcoma cell line was obtained from the European Collection of Authenticated Cell Cultures and cultured in DMEM/F12 medium (Life Technologies) supplemented with 10% fetal calf serum (Dutscher) and 4 μg/mL gentamicin.
To generate HEMC-SS spheroids, cells were harvested from culture flasks by trypsinization and seeded in 96-well non-adherent plates (Nunc) with 100 μL of 0.5% methylcellulose (Bio-Techne) diluted in the corresponding medium at a density of 1000 cells per well. Cultures were maintained in a 5% CO₂ humidified environment at 37°C.
Spheroid cell numbers were determined at different times (Day 1 to Day 10) after cell dissociation by incubation with Trypsin-EDTA (Life Technologies) for 5 minutes at 37°C.

Human HEMC-SS CHS cell line was obtained from the European Collection of Authenticated Cell Cultures, and cultured in DMEM/F12 medium (Life Technologies, Carlsab, CA, USA) supplemented with 10% fetal calf serum (Dutscher) and 4 μg/mL gentamicin.
To generate HEMC-SS spheroids, cells were harvested from culture flasks by trypsinization, and seeded in 96-well non-adherent plates (Nunc) with 100 μL of 0.5% methylcellulose (Bio-Techne) diluted in the whole culture medium at a density of 1000 cells per well. Cultures were maintained in a 5% CO₂ humidified environment at 37°C.
Spheroid doubling time was determined from exponential linear regression of cell counts from spheroids dissociated by incubation with trypsin-EDTA (Life Technologies) for 5 minutes at 37°C