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Related Concept Videos

Anatomy of the Intestines01:23

Anatomy of the Intestines

Although digestion of proteins, carbohydrates, and lipids may begin in the stomach, it is completed in the intestine. The absorption of nutrients, water, and electrolytes from food and drink also occurs in the intestine. The intestines can be divided into two structurally distinct organs—the small and large intestines.
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Inflammatory Bowel Disease III: Diagnostic Studies and Management I-Nutritional Therapy01:30

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Various diagnostic tests are employed in the diagnostic process for Inflammatory Bowel Disease (IBD), particularly to differentiate between Crohn's disease and ulcerative colitis.
Diagnostic studies
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Imaging Studies III: Gastrointestinal Motility Studies and Virtual Colonoscopy01:26

Imaging Studies III: Gastrointestinal Motility Studies and Virtual Colonoscopy

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Endoscopic Procedures II: Colonoscopy

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Endoscopic Procedures IV: Sigmoidoscopy and Laproscopy01:26

Endoscopic Procedures IV: Sigmoidoscopy and Laproscopy

Sigmoidoscopy and laparoscopy are distinct medical procedures that enable physicians to internally inspect different parts of the GI tract. Although they serve different purposes, each is essential for diagnosing and, in some cases, treating various medical conditions.
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Intestinal Obstruction I: Introduction

Intestinal obstruction is a partial or complete blockage of the small or large intestine that disrupts the normal flow of intestinal contents through the lumen. This interruption impairs digestion, absorption, and fluid balance, and may lead to serious complications if not treated promptly.Mechanical ObstructionMechanical obstruction occurs when a physical blockage prevents intestinal contents from passing, arising from within the lumen or the bowel wall, or from external compression.Adhesions,...

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The Ex Vivo Culture and Pattern Recognition Receptor Stimulation of Mouse Intestinal Organoids
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A Practical Guide to Developing and Troubleshooting Patient-Derived "Mini-Gut" Colorectal Organoids for Clinical

Rex Devasahayam Arokia Balaya1,2, Zahra Heydari1, Gobinda Sarkar1

  • 1Department of Gastroenterology and Hepatology, Mayo Clinic, Rochester, MN 55905, USA.

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|October 28, 2025
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Summary

This study details a protocol for generating patient-derived organoids (PDOs) from colorectal tissues. These PDOs are crucial for personalized medicine, drug screening, and understanding cancer mechanisms.

Keywords:
colorectal cancerpatient-derived organoidsthree-dimensional culture (3D)

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Area of Science:

  • Biotechnology
  • Gastroenterology
  • Oncology

Background:

  • Patient-derived organoids (PDOs) are valuable models for personalized medicine and cancer research.
  • PDOs replicate tumor heterogeneity, aiding drug screening and precision treatment strategies.
  • Understanding disease mechanisms and ex vivo exposures is enhanced by organoid models.

Purpose of the Study:

  • To present a detailed step-by-step protocol for generating organoids from normal, polyp, and tumor colorectal tissues.
  • To provide methods for tissue processing, crypt isolation, culture establishment, and polarity transition.
  • To include immunofluorescence staining and quality control for cellular characterization.

Main Methods:

  • Standardized protocol for colorectal tissue processing and crypt isolation.
  • Organoid culture establishment and basolateral to apical-out polarity transition.
  • Immunofluorescence staining for cellular characterization and quality control.

Main Results:

  • High-efficiency and reproducible generation of organoids from diverse colorectal tissues.
  • Successful implementation of methods for tissue processing, crypt isolation, and culture.
  • Standardized approach addresses technical challenges and improves success rates.

Conclusions:

  • The developed protocol enhances reproducibility in organoid research.
  • Organoid utility is expanded for translational applications in personalized medicine.
  • This resource is particularly valuable for colorectal cancer research and precision treatment.