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Recipient colon preoperative treatment with type I collagenase and fibronectin promotes the growth of transplanted enteric neural crest cells into Auerbach’s plexus

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Abstract

Purpose

Cell-based therapy is a potential treatment option for neurointestinal diseases by serving as a source of neural progenitor cells to replace missing or abnormal enteric neurons. Using an ex vivo transplantation model, we recently demonstrated that treatment with collagenase and fibronectin promotes infiltration of transplanted enteric neural crest cells (ENCCs) toward the colon lumen. The aim of this study was to determine whether this new method also promotes colonization of transplanted ENCCs in vivo.

Methods

Collagenase was applied locally on the anti-mesenteric area of the recipient colon using filter paper, followed by fibronectin. Neurospheres were generated from ENCCs isolated from fetal mouse intestines and transplanted into the collagenase and fibronectin-treated colon. Engraftment of neurospheres was confirmed by immunofluorescence.

Results

Neurospheres transplanted onto PBS- or fibronectin-treated colons were not observed to infiltrate to the muscle layer. However, when used in combination with type I collagenase and fibronectin in the recipient colon, transplanted neurospheres reached Auerbach’s plexus.

Conclusion

We demonstrated that transplanted neurospheres grow into Auerbach’s plexus in the recipient colon pretreated with collagenase and fibronectin.

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Acknowledgements

We thank Dr. Hideki Enomoto (Kobe University) for provision of Ednrb-hKikGR mice and Dr. Mamoru Tanida (Kanazawa Medical University) for helpful suggestions. We also thank Mitchell Arico from Edanz (https://jp.edanz.com/ac) for editing a draft of this manuscript. This work was supported in part by Innovation in Cell Therapy in the Hokuriku District of Japan through the Private University Branding Project of MEXT (RP2017-11), Collaborative Research from Kanazawa Medical University (C2016-3 and C2017-1), and JSPS KAKENHI (19K22669 and 22K06050).

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Author notes

  1. Tsuyoshi Kuwahara and Yoshitomo Yasui have contributed equally to this work.

Authors and Affiliations

  1. Department of Pediatric Surgery, Kanazawa Medical University, 1-1 Daigaku, Uchinada, Kahoku-gun, Ishikawa, 920-0293, Japan

    Tsuyoshi Kuwahara, Yoshitomo Yasui, Hisayoshi Yoshizaki, Miyuki Kohno & Hideaki Okajima

  2. Department of Pediatrics, Kanazawa Medical University, 1-1 Daigaku, Uchinada, Kahoku-gun, Ishikawa, 920-0293, Japan

    Mari Morikawa

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  1. Tsuyoshi Kuwahara
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  5. Miyuki Kohno
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Contributions

T.K., Y.Y. and H.Y. designed the studies and wrote the manuscript. T.K., Y.Y. and M.M. designed experiments, conducted in vivo studies, and analyzed data. H.Y., M.K. and H.O. supervised the project and edited the manuscript. All authors reviewed the manuscript.

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Correspondence to Hisayoshi Yoshizaki.

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Kuwahara, T., Yasui, Y., Yoshizaki, H. et al. Recipient colon preoperative treatment with type I collagenase and fibronectin promotes the growth of transplanted enteric neural crest cells into Auerbach’s plexus. Pediatr Surg Int 38, 1793–1798 (2022). https://doi.org/10.1007/s00383-022-05224-w

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