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Purpose <p>This study aimed to generate a hybrid colon lined with stratified esophageal squamous epithelium by replacing native colonic epithelium with esophageal organoids in mice.</p> Methods <p>Esophageal organoids were cultured from 8 to 10-week-old enhanced green fluorescent protein (EGFP) transgenic mice for 7–14 days before engraftment into syngeneic wild-type mice. A ~ 1.5&#xa0;cm proximal colon segment was chemically de-epithelialized using ethylenediaminetetraacetic acid and flushed with phosphate buffered saline. The organoid suspension was then introduced into the lumen. Target tissues were collected on postoperative days 2, 11, 14, or 28 (<i>n</i> = 5, respectively) and analyzed by Hematoxylin-Eosin staining and immunofluorescence.</p> Results <p>Cultured esophageal organoids were lined by stratified epithelial structures with Cytokeratin (CK)14 + basal cells and CK13 + luminal cells, resembling native esophageal epithelium. Following transplantation, EGFP + cells that adhered to the luminal surface of the colon, first developed an immature multilayered epithelium and then formed a mature, differentiated epithelium by day 14. Immunofluorescence confirmed the presence of spatially organized CK14 and CK13 expression and the absence of the colon-specific marker CK20, indicating preservation of esophageal identity in the graft.</p> Conclusions <p>Engraftment of esophageal organoids into the de-epithelialized colon resulted in the successful reconstitution of esophageal epithelium, preserving their original phenotype.</p>

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Regenerative reconstruction of mature esophageal epithelium by engraftment of esophageal organoids in de-epithelialized mouse colon

  • 期刊论文

  • 发表日期：

    2025年11月14日

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  • Xuxuan Rao,
  • Kazuto Suda,
  • Yuka Matsumoto,
  • Hiroyuki Koga,
  • Go Miyano,
  • Tetsuya Nakamura,
  • Atsuyuki Yamataka

摘要

Purpose

This study aimed to generate a hybrid colon lined with stratified esophageal squamous epithelium by replacing native colonic epithelium with esophageal organoids in mice.

Methods

Esophageal organoids were cultured from 8 to 10-week-old enhanced green fluorescent protein (EGFP) transgenic mice for 7–14 days before engraftment into syngeneic wild-type mice. A ~ 1.5 cm proximal colon segment was chemically de-epithelialized using ethylenediaminetetraacetic acid and flushed with phosphate buffered saline. The organoid suspension was then introduced into the lumen. Target tissues were collected on postoperative days 2, 11, 14, or 28 (n = 5, respectively) and analyzed by Hematoxylin-Eosin staining and immunofluorescence.

Results

Cultured esophageal organoids were lined by stratified epithelial structures with Cytokeratin (CK)14 + basal cells and CK13 + luminal cells, resembling native esophageal epithelium. Following transplantation, EGFP + cells that adhered to the luminal surface of the colon, first developed an immature multilayered epithelium and then formed a mature, differentiated epithelium by day 14. Immunofluorescence confirmed the presence of spatially organized CK14 and CK13 expression and the absence of the colon-specific marker CK20, indicating preservation of esophageal identity in the graft.

Conclusions

Engraftment of esophageal organoids into the de-epithelialized colon resulted in the successful reconstitution of esophageal epithelium, preserving their original phenotype.