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Updated: Jan 27, 2026

Generation of Apical-Out Intestinal Organoids and Assessment of Organoid Proliferation Rate Using EdU-Labeling
Published on: March 28, 2025
Apical constriction is necessary for crypt formation in small intestinal organoids
Leonie Hartl1, Guizela Huelsz-Prince1, Jeroen van Zon1
1AMOLF, Science Park 104, 1098 XG Amsterdam, the Netherlands.
Changes in cell shape, not just proliferation, drive the formation of crypts in small intestinal organoids. Myosin II activity causes cells to constrict, initiating bud formation for crypt development.
Area of Science:
- Gastroenterology
- Developmental Biology
- Tissue Engineering
Background:
- Small intestinal organoids are vital models for studying crypt homeostasis, cell fate, and biomechanics.
- Mechanisms of crypt budding from smooth organoid epithelium are not fully understood.
- Previous hypotheses suggested proliferation-induced tissue buckling initiates crypts.
Purpose of the Study:
- To investigate the mechanisms driving crypt budding in small intestinal organoids.
- To elucidate the role of cell morphology and molecular activity in crypt formation.
Main Methods:
- Observation of crypt formation in small intestinal organoids.
- Analysis of cell morphology changes during crypt initiation.
- Assessment of myosin II activity and its role in apical constriction.
Main Results:
- Crypt formation is preceded by local epithelial thickening and apicobasal cell elongation.
- Cells adopt a wedge-like shape with apical narrowing before evagination.
- Myosin II activity correlates with apical constriction, and its inhibition prevents bud formation.
Conclusions:
- Cellular morphology changes, specifically apical constriction, are crucial for crypt initiation in small intestinal organoids.
- Myosin II-driven apical constriction is a key force initiating bud formation.
- This study reveals a novel mechanism for crypt development in organoid models.
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