Extended Data Fig. 1: Single-cell-derived bladder organoids were maintained for over a year by short-term serial passaging or long-term culture.
From: Creation of bladder assembloids mimicking tissue regeneration and cancer
a, Schematic diagram illustrating the experimental strategies for the short- and long-term culture of mouse normal bladder organoids. b, Time-course images showing that single urothelial cells successfully generated a bladder organoid. Scale bar, 100 μm. c, Quantification of organoid size when cultured for 9 days at each passage. d, Quantification of the organoid-forming efficiency at each passage. e, Bright-field images of serially passaged short-term (day 9) bladder organoids from passage 1 to 20. Scale bars, 100 μm. f, Representative images of serially passaged short-term (day 18) bladder organoids analysed by bright-field images and immunostaining. Scale bars, 100 μm. g, Average size of long-term cultured bladder organoids at indicated day. h, Representative images of bladder organoids at indicated times during long-term culture, analysed by bright-field images and immunostaining. Scale bars, 100 μm. i, Short-term (day 14) and long-term (day 81) organoids compared with embryonic (E16) bladders and adult (p8week) bladders. Dotted lines demarcate the border between epithelium and stroma (white arrowheads, umbrella cells). Scale bars, 20 μm. j, The relative expression of Upk2, Upk1a, Foxa1, and Gata3 in short-term (day 14) organoids, as compared to long-term (day 81) organoids. Data are mean ± s.e.m. For details on statistics, sample sizes (n), and numbers of replications, see ‘Statistics and reproducibility’ (Methods).
