Figure 1.

Reprogramed CLiPs form 3D cysts on gelatin-coated dishes (see also ). (a) Standardized protocol for inducing the formation of three-dimensional (3D) cysts from cultured CLiPs on gelatin-coated dishes; CLiPs were pipetted three times on each of the first three days, and culture medium change were replaced on days 4, 7, and 10. (b) Phase contrast images at indicated time points showing enlargement of 3D cysts; scale bar = 100 μm. (c) Changes in cyst diameters at indicted time points; data are shown as the means ± standard deviations (s.d.; n ≥ 6), and more than 30 cysts were chosen for each experiment at all time points. (d) Immunostaining images of F-actin microfilaments in 3D cyst structures (d1) with hollow lumens (d2); scale bar = 100 μm. (e) Phase contrast images of 3D cysts derived from CLiPs at various passages; scale bar = 200 μm.