MiaPaCa-2 and Panc-1 cells (1,000 cells/well) were plated in 96-well extremely low attachment plates (Corning, Inc.) and treated with oxaliplatin (5 µM)/cisplatin (5 µM)/carboplatin (4 µM) at 37°C for 48 h. Fibroblast growth factor (20 ng/ml), human recombinant epidermal growth factor (20 ng/ml) and serum substitute B27 (1X) were mixed into serum-free DMEM/F-12 and were added to each well (all from Invitrogen; Thermo Fisher Scientific, Inc.). The number of spheres >50 mm in diameter was counted under a light microscope after 2 weeks of incubation at 37°C and 5% CO2.
Protocol detail
Chemotherapeutics Affect Pancreatic Cancer Spheres
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Carboplatin
Cells
Cisplatin
Epidermal growth factor
Fibroblast growth factor
Human
Microscope light
Oxaliplatin
Serum
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Variable analysis
independent variables
- Oxaliplatin (5 µM)
- Cisplatin (5 µM)
- Carboplatin (4 µM)
- Number of spheres > 50 mm in diameter
- Cell line (MiaPaCa-2 and Panc-1 cells)
- Cell density (1,000 cells/well)
- Plate type (96-well extremely low attachment plates)
- Incubation time (48 h)
- Incubation temperature (37°C)
- Growth factors (fibroblast growth factor (20 ng/ml), human recombinant epidermal growth factor (20 ng/ml), serum substitute B27 (1X))
- Culture medium (serum-free DMEM/F-12)
- No treatment control (not explicitly mentioned)
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