Design of a Device for Measuring Intestinal Epithelial Cell Migration Chris Shen BME 273 Oral Presentation #3 Advisor: D. Brent Polk, M.D.

Background on Cell Migration Driving force for cell movement provided by dynamic reorganization of actin cytoskeleton with protrusion at the front of the cell and retraction at the rear Lamellipodia and filopodia found at the cell’s leading edge provide forward movement of the rear of the cell During wound healing, cells move in unidirectional and synchronous manner towards damaged area

Preparation of Cell Samples Fibronectin is a plasma-derived protein commonly used as an attachment factor for the culture of many cell types under reduced or serum-free conditions. Dishes coated overnight with fibronectin to form cellular matrix upon which endothelial cells can migrate

Drill Tip Design Silicone tubing Pencil casing provides structural support Silicone dipped in methanol/dry ice mixture in order to sharpen to a fine point with razor blade

Equipment Setup Wound is made while drill is rotating ~60RPM User holds down lever all the way for 2-3 seconds and releases

Wound Identification 1.5million YAMC cells plated on 35mm dishes 2 3 4 5 6 7 8 1.5million YAMC cells plated on 35mm dishes Ink pen used to divide dish into numbered sections to locate wound under microscope

Wound Reproducibility

Comparison of Treatments No treatment EGF 0 hrs 8 hrs 24 hrs

Graphic Comparison

Future Work Giemsa staining to ensure underlying fibronectin matrix intact upon making wound Develop wound closure curve by taking multiple early time points (0-8hrs) Confirm effects of chemotactic factors through paired t-tests.