Fig. 6

Time course of rapamycin-induced cell detachment and EB formation. iPSCs were cultured in the presence of 200 nM rapamycin. Bright-field microscopic images were taken as untreated control (a) and after 1 day (b), 2 days (c), 3 days (d) and 6 days (e, f) of treatment. EB-like spheres were spontaneously formed after 6 days of rapamycin treatment. (c, e, f) The border of the iPSC colonies formed a line after day 6 to limit colony expansion sideways (arrowheads). (e) Areas of iPSC detachment were filled with a thin layer of iPSCs (*). (f) Four EB-like spheres of comparable size were captured in the fields (arrowheads). (g) Magnified view (4×) of the border area with arrowheads in f. Bar = 100 μm a–d; 200 μm e, f