Fig. 1: Isolation and culture of patient-derived organoids (PDOs) from non-muscle invasive bladder cancer (NMIBC) and muscle invasive bladder cancer (MIBC). | Nature Communications

Fig. 1: Isolation and culture of patient-derived organoids (PDOs) from non-muscle invasive bladder cancer (NMIBC) and muscle invasive bladder cancer (MIBC).

From: Bladder cancer organoids as a functional system to model different disease stages and therapy response

Fig. 1: Isolation and culture of patient-derived organoids (PDOs) from non-muscle invasive bladder cancer (NMIBC) and muscle invasive bladder cancer (MIBC).

a Scheme of the experimental protocol for bladder cancer (BLCa) organoids derivation and culture. Created with BioRender.com. b Number of PDO formation and no PDO formation over the total samples cultured for NMIBC (n = 24 biological samples) and MIBC (n = 25 biological samples). c, d Morphology of parental tumor (PTs, Hematoxylin and Eosin staining) and matched PDOs at passage (p) 1 (brightfield image, upper) for two representative cases (BLCa34, transurethral resection of bladder tumor, Ta stage (c); BLCa40, cystectomy, T2b stage (d)). Immunohistochemistry for Ki67 for PT, and whole-mount immunofluorescence staining for Ki67 for PDOs (bottom). e Viability assay of PDOs at p2 derived from 5 NMIBC and 5 MIBC samples after 96 h in culture. Each data point corresponds to one technical replicate (mean ± SD) at 96 h (normalized to the time 0 h) for one experiment (n represents the technical replicates: n = 10 for BLCa30 (96 h); n = 5 for BLCa22 (0 h), BLCa26 (0 h), BLCa30 (0 h), BLCa40 (0 h), BLCa51 (0 h), BLCa53 (0 h), and BLCa77 (0 h); n = 8 for BLCa34 (96 h), BLCa51 (96 h), BLCa22 (96 h), BLCa26 (96 h) and BLCa53 (96 h); n = 7 for BLCa40 (96 h), and BLCa86 (0 h); n = 6 for BLCa77 (96 h); n = 4 for BLCa60 (0 h and 96 h), BLCa86 (96 h), and BLCa34 (0 h)). Statistically significance between time 0 and 96 h was calculated by two-sided Welch’s test. f Fraction of cell types in PT/PDOs (p1) pairs for three representative cases (BLCa77 NMIBC, BLCa86 MIBC, and BLCa98 MIBC). g UMAP plot of cells derived from PT/PDOs (p1) pairs clustered by cell types. h Fraction of cells in cell cycle phases in PT/PDOs pairs. i, j Proportion of epithelial cells that correspond to each molecular class for the BLCa77 sample (i, UROMOL2021 classifier) and for BLCa86 and BLCa98 samples (j, Consensus classifier). Ba_Sq basal/squamous, LumNS luminal nonspecified, LumP luminal papillary, LumU luminal unstable, NE-like neuroendocrine-like, ns not significant.

Back to article page