Individual cells were suspended into medium. Each group was inoculated with l0 ml culture medium with 200 cells per dish, and the cells were evenly dispersed by gentle shaking. Routine culture for 3 weeks. We eliminated the medium once there was clone seen within the petri dish, followed by 15 min of 4% paraformaldehyde fixation, 10 min of 0.1% crystal violet staining, PBS washing, and the number of clones was counted.

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