Fig. 2.
CD34+ CSCs support IESCs proliferation. (A) Bright field microscopy analysis (Left) and percentage of organoids versus spheroids (Right) among intestinal structures grown in the indicated conditions for 5 d. Arrows indicate budding crypts, and an asterisk indicates organoid lumen; n = 4 from three independent experiments. (B) Confocal analysis of Ki-67 (red) expression in intestinal structures grown as described in A. (C) Intracellular FACS analysis of Ki-67 in organoids grown as indicated. n = 6 from two independent experiments. (D) Confocal analysis (Left) and quantification (Right) of WGA expression (green) in intestinal structures in indicated conditions. n = 8 from two independent experiments. (E) Percentage of GFP+ cells, as measured by FACS, in organoids harvested from Lgr5-egfp mice and grown as indicated; n = 6 from two independent experiments. (F) FACS analysis of GFP reporting Lgr5 expression in intestinal structures obtained from Lgr5-egfp mice and grown as indicated; n = 6 from two independent experiments. In A and C–F, values are mean ± SD. (Scale bars: 100 μm.) DAPI stains nuclei. ****P < 0.0001, ***P < 0.001, **P < 0.005, *P < 0.05.