1. TNF  Receptor Regulation of Intestinal Epithelial Cell Migration Chris Shen BME 273 Oral Presentation #1 Advisor: D. Brent Polk, M.D.

2. Background Tumor necrosis factor alpha (TNF  ) is a prominent cytokine in the pathogenesis of inflammatory bowel disease Effects on the intestinal epithelium remain poorly understood TNF  exerts its effects through two glycoprotein receptors on the cell membrane (TNF  -R1 and TNF  -R2)

3. Objectives Design a device and/or method to make reproducible wound models in a cultured cell monolayer Monitor wound closure via time-lapse video microscopy Use statistical analysis to determine differences in cell migration across varying conditions

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

5. Previous Work Literature research on cell migration and mechanism of TNF  intracellular signaling Discussion with specialists: Dr. Brent Polk Dr. Wei Tong Dr. Jim McKanna

6. Current Work Development of drill tip Silicone tubing Sewing needle Fine drill bit Imaging software (calculating surface area) Mouse pen Function within program Potential applications with Free Electron Laser

7. Future Work Plot concentration curve to establish levels of TNF  inhibiting/promoting proliferation Determination of peak time to observe greatest statistical difference across various treatments Evaluation of TNF  R1 and R2 receptors