Colonoscopic Guided Murine Wound Model: A Method to Isolate Biopsy Induced Wound Bed from Colon to Understand the Healing Process

Abstract

Source: Montrose, D. C., et al. Performing Colonoscopic-Guided Pinch Biopsies in Mice and Evaluating Subsequent Tissue Changes. J. Vis. Exp. (2021).

In this video, we describe a procedure to isolate the colonoscopic-induced wound bed at an appropriate time point for detailed molecular analysis. This analysis helps to understand the wound healing process in inflammatory conditions of the GI tract.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Collection of the wound bed for molecular analysis Euthanize mouse using CO2 asphyxiation followed by cervical dislocation (or equivalent technique) on the selected day following biopsy. Harvest the distal region of the colon by opening the skin and abdominal muscle to expose the body cavity. Place closed sciss…

Representative Results

Figure 1: Ex vivo and sectioned images of wound bed. (A) A colon was harvested from a mouse 2 days following biopsy, stained with 0.2% methylene blue and imaged under a dissecting microscope. The dashed circle indicates the edges of the wound bed. The black line indicates the proper location to bisect the wound bed/colon prior to embedding for sectioning. (B) Representative image of an H&E-st…

Materials

Biopsy forceps, 3 Fr	Karl Storz	61071ZJ
isofluorane	Covetrus	2905
methylene blue	Sigma-Aldrich	M9140
micro iris scissors	Integra	18-1619
scalpal with #10 blade	Hill-Rom	372610