TNF Receptor Regulation of Intestinal Epithelial Cell

Migration

Chris Shen

BME 273 Oral Presentation #1

Advisor: D. Brent Polk, M.D.

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)

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

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

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

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

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