Single-cell suspensions were cultured in MammoCult Human MediumKit (Stemcell Technologies, Cambridge, MA, USA) at a density of 5,000 cells per well of a 6-well ultralow attachment culture plate (Corning) for 10 days as described [27 (link)]. Tumorspheres with a diameter >50 microns were counted under an inverted microscope in triplicate wells.
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