Fig. 3

Characterization of transduced RN5-EOS-Puro2 cells and confirmation of properties of stemness. a Highly enriched MSC were obtained after 2 weeks of selection with puromycin; note the increase in the GFP-positive population over time reaching 63.4% after 14 days (right lower image). b Quantification of the MSC GFP⁺ population determined by flow cytometry from the beginning of puromycin selection (D0) up to D21. c Sphere-formation assay: highly enriched MSC from the above selection were cultured in ultra-low adherent 24-well plates and spheres were formed in approximately 7–10 days. d Highly enriched MSC were stained for Oct4, which was positive in the nucleus (oil immersion objective; 60×)