Design of a Device for Measuring Intestinal

Epithelial Cell Migration

Chris Shen

BME 273 Oral Presentation #2

Advisor: D. Brent Polk, M.D.

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.

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)

Project Requirements

Assay in preparing cell samples

Physical device to make wounds

Wound analysis

Comparative analysis

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

Previous Work Assay for setting up cell samples completed

Coat dishes overnight with matrixSerum starve YAMC cells 16hrs in .5%

media Preliminary device developed

Drill pressStandard drillCustom-designed tip

Initial Assessment of Device Performance

Wound shape relatively circular

Noticeable wound closure after 7hrs

Calculation of surface areaAutomatedManual

T=0hrs

T=7hrs

Problems Encountered

Noticeable damage to cell matrix

Rough tip created internal irregularities within wounds

Current Work

Improve quality of device tip

Development of assay for staining cellular

matrix

Determine optimum method for measuring

wound surface area

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