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Sphere formation cultures.
Cells (1 × 103) were seeded into each well of ultra-low attachment 6-well plates (Corning, NY, USA) and cultured in DMEM supplemented with 10% (v/v) FBS at 37 °C under 5% CO2. Phase-contrast photomicrographs of cell masses derived from single cells; 1 × 103 HSC3, HSC4, Ca9-22, KB, DU145 and SAS cells were grown on ultra-low attachment 6-well plates.
