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. 2023 Aug 31;12(17):2188.
doi: 10.3390/cells12172188.

A Protocol for Organoids from the Urine of Bladder Cancer Patients

Simon Walz  1 , Paul Pollehne  2 , Ruizhi Geng  2 , Johannes Schneider  2 , Moritz Maas  1 , Wilhelm K Aicher  2 , Arnulf Stenzl  1 , Bastian Amend  1 , Niklas Harland  1
Affiliations

A Protocol for Organoids from the Urine of Bladder Cancer Patients

Simon Walz et al. Cells. .

Abstract

This study investigates the feasibility of establishing urine-derived tumor organoids from bladder cancer (BC) patients as an alternative to tissue-derived organoids. BC is one of the most common cancers worldwide and current diagnostic methods involve invasive procedures. Here, we investigated the potential of using urine samples, which contain exfoliated tumor cells, to generate urine-derived BC organoids (uBCOs). Urine samples from 29 BC patients were collected and cells were isolated and cultured in a three-dimensional matrix. The establishment and primary expansion of uBCOs were successful in 83% of the specimens investigated. The culturing efficiency of uBCOs was comparable to cancer tissue-derived organoids. Immunohistochemistry and immunofluorescence to characterize the uBCOs exhibited similar expressions of BC markers compared to the parental tumor. These findings suggest that urine-derived BC organoids hold promise as a non-invasive tool for studying BC and evaluating therapeutic responses. This approach could potentially minimize the need for invasive procedures and provide a platform for personalized drug screening. Further research in this area may lead to improved diagnostic and treatment strategies for BC patients.

Keywords: bladder cancer; organoids; personalized medicine.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Method and workflow of the culturing process of uBCOs. The materials and methods section contains a comprehensive and detailed step-by-step workflow, along with the materials and reagents used in the study. Abbreviations: BC, bladder cancer; uBCO, urine-derived bladder cancer organoid. Created with biorender.com (accessed 12 May 2023).
Figure 2
Figure 2
Culturing efficiency of uBCOs (indicated in black) and tBCOs (indicated in grey hatched) (A) with respect to the individual passages (efficiency rate). Culturing efficiency according to patient characteristics and histopathological features with regard to the maximum achieved passage (B–I). Dots represent data from individual organoids. Boxes indicate median and 25th and 75th percentiles with min/max whiskers. p-values indicate differences between two groups (A: two-sided Fischer exact test; B: Mann–Whitney-U test). The ρ-values indicate linear correlations between two parameters (Spearman correlation). Abbreviations: uBCO, urine-derived bladder cancer organoid; tBCO, tissue-derived bladder cancer organoid; NMIBC, non-muscle- invasive bladder cancer; MIBC, muscle-invasive bladder cancer.
Figure 3
Figure 3
Light microscopic images of uBCO #027 during cultivation in basal membrane extract in the upper row on the first, fourth, and thirteenth day of the first passage, in the middle and lower row on the first and seventh day of the third and fourth passage, respectively. Size bars indicate 100 mm. Abbreviations: uBCO, urine-derived bladder cancer organoid; P, passage; D, day.
Figure 4
Figure 4
Immunohistochemical images from parental primary BC (left column) and autologous, uBCO #027 (middle column) in the fifth passage. The left column displays fluorescence staining of relevant antigens in urothelial cancer or tumor stem cell antigens in the corresponding uBCO #027 (passage 5). The tBCO only reached the first passage and could, therefore, not be implemented in the analysis. Abbreviations: BC, bladder cancer; uBCO, urine-derived bladder cancer organoid; tBCO, tissue-derived bladder cancer organoid HE, hematoxylin-eosin; GATA, glutamyl amino tranverase A; CK, cytoceratine; p, protein; CD, cluster of differentiation.
See this image and copyright information in PMC

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