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In vitro investigation of the differentiation of enteric neural crest-derived cells following transplantation of aganglionic gut in a mouse model

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Abstract

Purpose

Cell therapy is a promising approach to treat enteric neuropathies such as Hirschsprung disease (HD). Recent studies have reported that enteric neurons derived from stem cells (ENCCs) can be grafted into the HD colon. Thus, we investigated the migration and generation of enteric neurospheres from SOX10-VENUS+ mice after transplantation into control or Ednrb KO mice, which are a model of HD.

Methods

Single-cell suspensions were isolated from the fetal guts of SOX10-VENUS+ mice E13.5 and dissociated. These cells were cultured for 7 days under non-adherent conditions to generate neurospheres, which were co-cultured with dissociated control or SOX10-VENUS− Ednrb KO mouse gut on E13.5. 4 days later, these cells were fixed and the expression of the neuronal marker, Tuj1, was evaluated.

Results

Transplanted neurospheres had undergone abundant neuronal migration and differentiation of ENCCs in the control gut compared with the HD gut. The average length and intersections were significantly decreased in HD colon compared with controls (p < 0.05), and a similar pattern was observed in the HD small intestine (p < 0.05).

Conclusions

We demonstrated that transplanted ENCCs did not differentiate properly in HD gut. These results highlight the importance of the neuronal environment in the recipient gut for enteric nervous system development.

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Acknowledgements

The authors thank Ms Mirei Takahashi for excellent technical support.

Funding

This work was supported by JSPS KAKENHI Grant no. 19K09103.

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Authors and Affiliations

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

    Naho Fujiwara, Katsumi Miyahara, Nana Nakazawa-Tanaka & Atsuyuki Yamataka

  2. Department of Pediatric Surgery, Juntendo University Nerima Hospital, Nerima-ku, Tokyo, Japan

    Nana Nakazawa-Tanaka

  3. Intractable Disease Research Center, Juntendo University School of Medicine, Bunkyo-ku, Tokyo, Japan

    Chihiro Akazawa

Authors
  1. Naho Fujiwara
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  2. Katsumi Miyahara
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  3. Nana Nakazawa-Tanaka
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  4. Chihiro Akazawa
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  5. Atsuyuki Yamataka
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Correspondence to Naho Fujiwara.

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Fujiwara, N., Miyahara, K., Nakazawa-Tanaka, N. et al. In vitro investigation of the differentiation of enteric neural crest-derived cells following transplantation of aganglionic gut in a mouse model. Pediatr Surg Int 38, 755–759 (2022). https://doi.org/10.1007/s00383-022-05105-2

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  • DOI: https://doi.org/10.1007/s00383-022-05105-2

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