Fig. 2.

ISCs from aged mice exhibit significant deficiency in differentiation in subculture. a-c Cultured crypts were dissociated and passaged on day 14 after primary plating. a,b Absolute number (a) and percentage (b) of grown out structures. c Representative pictures of indicated groups on day 4 after secondary plating. Small cysts: diameters ≤70 μm; big cysts: diameters >70 μm; small organoids: with crypt-villus architectures, budding number ≤ 3; big organoids: with crypt-villus architectures, budding number > 3. Arrow heads indicate typical organoids; arrows indicate typical big cysts. Data are displayed as mean ± SEM. *, P < 0.05; ***, P < 0.001; ****, P < 0.0001. ns, not significant. Unpaired two tailed Student’s t test was used. Scale bar: 200 μm