A suspension of parental MDA-MB-231 cells was diluted using culture medium to a cell density of approximately 1 cell/0.1 μl. A droplet of 0.1 μl of cell suspension was placed in each well of a 96-well plate by pipetting followed by microscopy inspection to examine the number of cells in the deposited droplet. For wells containing a single cell, 200 μl of culture medium was subsequently added to allow for cell growth into SCCs. The culture medium was then replaced regularly every 3 to 4 days, and SCCs were subsequently transferred to 24-well plates, 6-well plates, and 10-cm petri dishes after they became confluent. SCCs were then frozen down and thawed for further experiments.

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