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

2. 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

3. 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

4. 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

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

6. Wound Identification 1.5million YAMC cells plated on 35mm dishes2 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

7. Wound Reproducibility

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

9. Graphic Comparison

10. 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.