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Successful engraftment of bladder organoids in de-epithelialized mouse colon

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Abstract

Purpose

To engraft bladder organoids (BO) on de-epithelialized mouse colon using an epithelial replacement technique.

Methods

BO cultured using bladder specimens from enhanced green fluorescent protein (EGFP) transgenic mice were engrafted to replace proximal colon epithelium stripped from an approximately 1 cm long target site in syngeneic wild-type recipient mice (n = 9) by exposure to ethylenediaminetetraacetic acid by infusion and flushing with phosphate buffered saline. Target sites were harvested on postoperative days 2, 7, and 28 for hematoxylin–eosin staining and immunofluorescence.

Results

Histology on postoperative days 7 and 28 showed BO derived EGFP + cells forming multiple layers on the luminal surface of the colon. Immunohistochemistry showed that EGFP + areas were positive for CK5 and CK14, markers for basal and immature subtype urothelium, respectively, but negative for CA2, a marker for colonic epithelium. Ki67 was detected predominantly in the basal parts of EGFP + areas on postoperative day 7 and day 28.

Conclusions

This is the first report of successful engraftment of BO in de-epithelialized colon with urothelial tissue reconstituted by actively proliferating cells. This technique could be developed for augmentation cystoplasty to prevent bladder calculi formation and malignant transformation.

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Data availability statement

Data are available on reasonable request from the authors.

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Acknowledgements

We thank the Laboratory of Morphology and Image Analysis and the Laboratory of Molecular and Biochemical Research, Research Support Center, Juntendo University Graduate School of Medicine for technical assistance. This study was supported by Japan Society for the Promotion of Science KAKENHI grants (20K21637 and 19K09083), GlaxoSmithKline Japan Research Grant 2019 (D-17), GlaxoSmithKline Japan Research Grant 2021 (AS2021A000149025), and Juntendo University Project Grant (No. 2020-19). Tetsuya Nakamura belongs to the Department of Research and Development for Organoids, which is endowed by the Japanese pharmaceutical group, Eisai Co., Ltd.

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Authors and Affiliations

  1. Department of Pediatric General and Urogenital Surgery, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-Ku, Tokyo, 113-8421, Japan

    Kazuto Suda, Yuka Matsumoto, Takanori Ochi, Hiroyuki Koga, Geoffrey J. Lane & Atsuyuki Yamataka

  2. Department of Research and Development for Organoids, Juntendo University Graduate School of Medicine, 2-1-1 Hongo, Bunkyo-Ku, Tokyo, 113-8421, Japan

    Kazuto Suda, Yuka Matsumoto, Hiroyuki Koga & Tetsuya Nakamura

  3. Department of Neurology, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-Ku, Tokyo, 113-8421, Japan

    Nobutaka Hattori

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  1. Kazuto Suda
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  2. Yuka Matsumoto
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  8. Atsuyuki Yamataka
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Contributions

KS and AY designed the study. TO, HK, NH, and TN provided the conceptual advice. KS, YM, and TN performed the organoid culture and engraftment experiments. KS, YM, TN, and AY analyzed the data. KS, GL, TN, and AY wrote the manuscript.

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Correspondence to Kazuto Suda.

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Suda, K., Matsumoto, Y., Ochi, T. et al. Successful engraftment of bladder organoids in de-epithelialized mouse colon. Pediatr Surg Int 39, 14 (2023). https://doi.org/10.1007/s00383-022-05294-w

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  • DOI: https://doi.org/10.1007/s00383-022-05294-w

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