BMSCs treated at a density of 2 × 103 were seeded in a 5-cm dish, and the medium was refreshed every 3 days. After 12 days of culture, the dish was washed using PBS twice and was fixed with 4% paraformaldehyde (Sigma-Aldrich Corp, St Louis, MO, USA) for 30 min. Then, 2% crystal violet (Amresco, Solon, OH, USA) was used to stain the clones. Clone that contains more than 50 cells under the stereomicroscope was defined as a colony.
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