1 Fibroblast Migration Increased by PEG-EGF in Hydrogels Student Name Date.

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1 Fibroblast Migration Increased by PEG-EGF in Hydrogels Student Name Date

2 Objective Does epidermal growth factor (EGF) tethered to PEG increase cell number and migration of fibroblasts in hydrogels?

3 Measuring Cell Proliferation Hydrogels with GGLG-PAGGK-PEG were prepared with or without PEG-EGF 20,000 cells placed on media-soaked gel and incubated 48 hours PCNA stained cells were counted Control performed similarly with liquid media and cell number determined with Coulter Counter

4 Measuring Cell Migration Hydrogels on membranes were prepared containing either PEG-EGF or PEG-RGDS, both PEG-EGF and PEG-RGDS, or neither 50,000 cells were added and incubated for 7 days Calculated migration index as % of cells that move through membrane as determined by Coulter Counter

5 EGF Increases Cell Number No EGF Number of cells EGF PEG-EGF - toxin + toxin More cells in presence of EGF Mean+/-s.d.; n=6 20,000 initial cell number * p=0.07 vs. no EGF

6 EGF Increases Number of Cells in S-Phase More cells in S-phase with EGF Mean+/-s.d.; n=6 * p=0.05 vs. no EGF No EGF Percentage of cells EGF PEG-EGF - toxin + toxin * *

7 EGF in Hydrogels EGF increases number of cells above 20,000 initial cell number (shown as dashed line) Mean+/-s.d.; n=7 * p=0.05 vs. no EGF No EGF EGF PEG-EGF * * 30,000 25,000 20,000 15,000 10,000 5, ,000 Number of cells

8 Migration occurs with RGDS and EGF+RGDS Migration occurs with RGDS and RGDS+EGF Migration doesn’t occur with EGF alone Migration occurs in presence of toxin PEG EGF RGDS Number of cells EGF & RGDS - toxin + toxin Mean +/- s.d. n=4

9 Fibroblast Migration Increased by PEG-EGF PEG-EGF increases cell number and percent of S-phase cells PEG-EGF with PEG-RGDS causes migration through hydrogel

10 Data and results taken from the paper Effects of Epidermal Growth Factor on Fibroblast Migration through Biomimetic Hydrogels, A.S. Gobin, J.L. West, Biotechnol. Prog. 2003, 19, Dr. A. Saterbak accepts all responsibility for any misrepresentation of any part of the paper. Poster developed entirely for pedagogical purpose for BIOE342.