Studying the Epithelial Effects of Intestinal Inflammation <em>In Vitro</em> on Established Murine Colonoids

Erin Crawford; Heather L. Mentrup; Elizabeth A. Novak; Kevin P. Mollen

doi:10.3791/64804

Journal

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Biology

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Studying the Epithelial Effects of Intestinal Inflammation In Vitro on Established Murine Colonoids

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Biology

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JoVE Journal Biology

Studying the Epithelial Effects of Intestinal Inflammation In Vitro on Established Murine Colonoids

Studying the Epithelial Effects of Intestinal Inflammation In Vitro on Established Murine Colonoids

897 Views

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06:31 min

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June 02, 2023

DOI:

10.3791/64804-v

DOI:

10.3791/64804-v

•

06:31 min

June 02, 2023

•

8 Views

Erin Crawford*¹, Heather L. Mentrup*^2,3, Elizabeth A. Novak^2,3, Kevin P. Mollen^2,3

¹Division of Pediatric Gastroenterology, Hepatology and Nutrition,UPMC Children’s Hospital of Pittsburgh, ²Division of Pediatric Surgery,UPMC Children’s Hospital of Pittsburgh, ³Department of Surgery,University of Pittsburgh School of Medicine

Transcript

Through our research, we aim to explore how murine colonoids could be modified from a conventional colonoid culture system, to study certain aspects of intestinal physiology and their alteration in the presence of an inflamed disease state. Studying barrier function and intestinal physiology in disease states like inflammatory bowel disease is limited when using a traditional murine colonoid culture system that reflects only stem cell physiology. Our protocol is advantageous in that it provides the reader with multiple techniques to study how inflammatory mediators affect the intestinal epithelium, by either terminally differentiating them, or creating a 2D monolayer system.

In the future, our laboratory will concentrate on comprehending how the human colonoid culture system can be altered to investigate different aspects of intestinal physiology and diseases, particularly inflammatory bowel disease.

Summary

We describe a protocol detailing the isolation of murine colonic crypts for the development of 3-dimensional colonoids. The established colonoids can then be terminally differentiated to reflect the cellular composition of the host epithelium prior to receiving an inflammatory challenge or being directed to establish an epithelial monolayer.