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Related Experiment Video

Updated: Jan 17, 2026

Three-Dimensional Culture of Murine Colonic Crypts to Study Intestinal Stem Cell Function Ex Vivo
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Exploring the Morphologic Characteristics of Intraperitoneally Injected Three-Dimensional-Cultured Tonsil-Derived

Ju Ran Byeon1, Eun Mi Song1, Yang Hee Joo1

  • 1Department of Internal Medicine, Ewha Medical Research Institute College of Medicine, Ewha Womans University, 260 Gonghangdae-ro, Gangseo-gu, Seoul 07804, Korea.

Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada
|September 24, 2025
PubMed
Summary

Three-dimensional tonsil-derived mesenchymal stem cells (3D-TMSCs) form aggregates with immune cells in mice with colitis. These structures show viability and paracrine effects, supporting their use in treating inflammatory bowel disease.

Keywords:
electron microscopyextracellular vesicleinflammatory bowel diseasemesenchymal stem cells

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Area of Science:

  • Regenerative Medicine
  • Immunology
  • Gastroenterology

Background:

  • Three-dimensional (3D)-cultured tonsil-derived mesenchymal stem cells (TMSCs) demonstrate significant therapeutic potential in preclinical models of colitis.
  • Understanding the in vivo behavior of these 3D-TMSCs is crucial for optimizing their therapeutic application.

Purpose of the Study:

  • To investigate the in vivo localization and formation of 3D-TMSCs within the peritoneal cavity of mice with dextran sulfate sodium-induced chronic colitis.
  • To characterize the interaction between 3D-TMSCs and host immune cells at the ultrastructural level.

Main Methods:

  • Mice with chronic colitis received intraperitoneal injections of 3D-TMSCs.
  • Immunofluorescence and electron microscopy were employed to assess the formation and localization of TMSC aggregates.
  • Human-origin TMSCs were identified using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) staining.

Main Results:

  • Spherical aggregates of 3D-TMSCs were observed in the peritoneal cavity, co-localizing with mouse immune cells.
  • Electron microscopy revealed extensive cell-to-cell connections, with immune cells covering the 3D-TMSCs via pseudopodia and microvilli.
  • Viability markers, including rough endoplasmic reticulum and mitochondria, along with autophagosomes and extracellular vesicles, were identified within the 3D-TMSC aggregates after 15 days.

Conclusions:

  • 3D-TMSCs form integrated structures with host immune cells in vivo, demonstrating sustained viability and paracrine activity.
  • These findings support the potential of 3D-TMSCs as an advanced therapeutic strategy for inflammatory bowel disease.