1. Design of a Device for Measuring Intestinal Epithelial Cell Migration Chris Shen BME 273 Oral Presentation #2 Advisor: D. Brent Polk, M.D.

2. Project Overview Design a device and/or method to make reproducible wound models in a cultured cell monolayer in order to study the effects of chemical factors which promote/inhibit intestinal cell migration.

3. Purpose The effects of various growth factors within the intestinal epithelium are poorly understood Various pathological conditions exist which affect rate of intestinal cell migration Crohn’s Disease Inflammatory Bowel Disease (IBD)

4. Project Requirements Assay in preparing cell samples Physical device to make wounds Wound analysis Comparative analysis

5. Device Specifications Produce 1~2mm diameter round wounds marked for identification to be photographed under standard light microscope No removal of underlying cellular matrix upon disrupting cell monolayer Performance of device relatively independent of the dexterity of the user

6. Previous Work Assay for setting up cell samples completed Coat dishes overnight with matrix Serum starve YAMC cells 16hrs in.5% media Preliminary device developed Drill press Standard drill Custom-designed tip

7. Initial Assessment of Device Performance Wound shape relatively circular Noticeable wound closure after 7hrs Calculation of surface area Automated Manual T=0hrs T=7hrs

8. Problems Encountered Noticeable damage to cell matrix Rough tip created internal irregularities within wounds

9. Current Work Improve quality of device tip Development of assay for staining cellular matrix Determine optimum method for measuring wound surface area

10. Future Work Determine time interval at which ~50% wound closure occurs under standard conditions Administer various chemical factors which stimulate/inhibit cell migration at known concentrations Evaluate efficacy of the procedure via comparative analysis