6 well ultra low attachment surface polystyrene culture plates

Human PDAC cell lines, AsPC‐1, Capan‐2, Panc‐1, and SW1990, were obtained from American Type Culture Collection. All cell lines were cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) with 10% FBS (MRC Biotechnology Co. Ltd.; cat. no. CCS30009.02) and 100 U/ml penicillin at 37°C in a humidified incubator with 95% air and 5% CO₂.
When the adherent cells were in the logarithmic growth stage, they were digested and resuspended, and the cell concentration was adjusted to 1×10⁴ cells/ml and inoculated in the complete MammoCult™ Human Medium (Stemcell Technologies, Inc.) using 6‐well ultra‐low attachment Surface Polystyrene culture plates (Corning, Inc.). After 10–14 days, we observed and imaged the morphology and quantity of pancreatic cell spheres using an inverted light microscope.

The human pancreatic adenocarcinoma cell lines AsPC-1, Capan-2, PANC-1, and SW1990 were obtained from the American Type Culture Collection (ATCC, Rockville, Md.). All cell lines were cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc. Waltham, Mass.) with 10% FBS (MRC Biotechnology Co. Ltd.; cat. no. CCS30009.02. Cincinnati, Ohio) and 100 U/ml penicillin at 37 °C in a humidified incubator with 95% air and 5% CO₂.
When the adherent PANC-1 cells were 90% confluent, they were digested and resuspended, and the concentration was adjusted to 1 × 10⁴ cells/ml. The cells were then inoculated into complete MammoCult™ Human Medium (Stemcell Technologies, Inc. Vancouver, Canada) using 6-well ultra-low attachment surface polystyrene culture plates (Corning. Inc. Corning, N.Y.). After 15 days, we observed and imaged the morphology and quantity of the pancreatic cell spheres using an inverted light microscope.