A Preclinical Model to Assess Intestinal Barrier Integrity Using Canine Enteroids and Colonoids

Area of Science:

• Cell Biology
• Biotechnology
• Gastroenterology

Background:

• Conventional 2D cell cultures like Caco-2 and MDCK cells have limitations in mimicking in vivo biology.
• Three-dimensional organoid technology presents a promising alternative for more accurate in vitro models.

Purpose of the Study:

• To compare the characteristics of canine intestinal organoids (enteroids, ENT, and colonoids, COL) with conventional 2D cell cultures (Caco-2, MDCK).
• To evaluate the microanatomy, monolayer integrity, and biological relevance of canine organoids versus 2D models.

Main Methods:

• Light and transmission electron microscopy were used to assess microanatomy.
• Transepithelial electrical resistance (TEER) measurements evaluated monolayer integrity.
• Comparison of ENT, COL, Caco-2, and MDCK cell systems.

Main Results:

• Canine ENT monolayers showed TEER values closer to human small intestine values than Caco-2 and MDCK.
• Canine ENT demonstrated superior monolayer stability compared to Caco-2 and MDCK cells.
• Canine ENT and COL exclusively produced mucus, unlike Caco-2 and MDCK.

Conclusions:

• Canine intestinal organoids represent a more biologically relevant in vitro model.
• Organoid systems address the inherent limitations of conventional 2D cell culture systems.
• This canine organoid model holds potential for advanced in vitro studies.