Human PDAC PANC−1 cells (ATCC-CRL−1469), human pancreas adenocarcinoma ascites metastasis cells AsPC−1 (ATCC-CRL−1682), and undifferentiated human pancreatic carcinoma MIAPaCa-2 cells (ATCC-CRL−1420) were purchased from ATCC (LGC Standard s.r.l., Sesto San Giovanni (MI), Italy), and were cultured as previously reported [42 (link)]. Human pancreatic cancer stllate cancer-associated fibroblasts (CAF08, PC00B5) were purchased from Vitro Biopharma (Golden, Colorado). All materials for cell culturing were purchased from EuroClone (Pero, Milan, Italy).
For PANC−1 tumor homo-spheroids formation, cells were seeded at a density of 5000 cells per well on 96-well Corning Spheroid Microplate (Corning, NY, USA) and incubated at 37 °C under 5% CO2 for 5 days before performing experiments. The spheroids of PANC−1 in co-culture with CAF cells were obtained, as described above for homospheroids using a 2:1 PANC−1:CAF ratio. Spheroid formation was assessed using OLYMPUS CKX41 microscope (OLYMPUS, Tokyo, Japan).
For PANC−1 tumor homo-spheroids formation, cells were seeded at a density of 5000 cells per well on 96-well Corning Spheroid Microplate (Corning, NY, USA) and incubated at 37 °C under 5% CO2 for 5 days before performing experiments. The spheroids of PANC−1 in co-culture with CAF cells were obtained, as described above for homospheroids using a 2:1 PANC−1:CAF ratio. Spheroid formation was assessed using OLYMPUS CKX41 microscope (OLYMPUS, Tokyo, Japan).
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