1. Development of 3D Breast Cancer
Models in PEG-Fibrinogen Hydrogels
Kelsey Henderson, Shantanu Pradhan, Dr. Elizabeth A Lipke
Department of Chemical Engineering
Auburn University, Auburn, AL, USA
July 24, 2015

2. Breast Cancer
• Most common cancer in
women
• Per year in the US: over
200,000 women diagnosed,
over 40,000 die
http://www.keep-a-breast.org/get-educated/cancer-facts/
http://keionline.org/node/1042
Ratio of deaths to new cases of
breast cancer in the United States

3. Breast Cancer Molecular Subtypes
http://labmed.ascpjournals.org/content/41/6/364.figures-only

4. 2D Cell Culture
Problem: Not accurate depiction of how cancer
cells grow in the body
https://www.spe
ctrumchemical.co
m/OA_HTML/lab-
supplies-
products_Corning
R-Sterile-Cell-
Culture-
Flasks_305276.js
p?section=23372
MCF-7 (ER+/PR+) SK-BR-3 (HER2+) MDA-MB-231 (TNBC)

5. 3D Cell Culture: Cells Encapsulated in Hydrogels
http://pubs.rsc.org/en/content/articl
ehtml/2013/cs/c3cs60040h

6. MCF-7 (ER+/PR+)
SK-BR-3 (HER2+)
MDA-MB-231 (TNBC)
Breast cancer cells encapsulated in hydrogels on day 6

7. Biomaterial for Cell Encapsulation
PEG-Fibrinogen:
• Cancer cells secret fibrinogen: promotes tumorigenesis
and angiogenic growth
• Poly(ethylene glycol) diacrylate provides stiffness and
architectural support for cell growth
• Imitates body tissue

8. Experimental Procedure
• Encapsulated all three cell lines (MCF-7,
MDA-MB-231, SK-BR-3) within PEG-Fb
hydrogels
• Phase contrast imaging at time points:
0, 3, 6, 9, 12, 15 days
• Measured diameter, circularity, area
• Immunostaining for phalloidin, Ki67,
vimentin, E-Cadherin, αvβ3 integrin, β1
integrin

9. MCF-7 progression of encapsulated cells
Day 0 Day 3 Day 15

10. Increase of Colony Area in 3D Culture
A B B C C C
MCF-7 (ER/PR+)
A B C C C C
SK-BR-3 (HER2+)
A B B B B B
MDA-MB-231 (TNBC)

11. Increase of Colony Area in 3D Culture
MCF-7 (ER+/PR+) day 6 SK-BR-3 (HER+) day 6 MDA-MB-231 (TNBC) day 6

12. Increase of Colony Diameter in 3D Culture
A B C D D D
MCF-7 (ER/PR+)
A B B B B B
MDA-MB-231 (TNBC)
A B C C C C
SK-BR-3 (HER2+)

13. Increase of Colony Diameter in 3D Culture
MCF-7 (ER+/PR+) day 6 SK-BR-3 (HER+) day 6 MDA-MB-231 (TNBC) day 6

14. Decrease of Colony Circularity in 3D Culture
A B BC BC CD D
MCF-7 (ER/PR+)
A B BC BC BC C
MDA-MB-231 (TNBC)
A B B B C C
SK-BR-3 (HER2+)

15. Decrease of Colony Circularity in 3D Culture
MCF-7 (ER+/PR+) day 6 SK-BR-3 (HER+) day 6 MDA-MB-231 (TNBC) day 6

16. Cell structure and morphology
Hoechst Phalloidin
MCF-7 (ER+/PR+) MDA-MB-231 (TNBC)SK-BR-3 (HER2+)
2-D3-D

17. Cell proliferation
Hoechst Ki67
2-D3-D
MCF-7 (ER+/PR+) SK-BR-3 (HER2+) MDA-MB-231 (TNBC)

18. Epithelial vs. Mesenchymal Cells

19. Mesenchymal cells express higher vimentin
Hoechst Vimentin
3-D2-D
MCF-7 (ER+/PR+) SK-BR-3 (HER2+) MDA-MB-231 (TNBC)

20. Epithelial cells express more E-Cadherin
Hoechst E-Cadherin
3-D
MCF-7 (ER+/PR+) SK-BR-3 (HER2+) MDA-MB-231 (TNBC)

21. MDA-MB-231 cells exhibit highest αvβ3
Hoechst αvβ3
3-D2-D
MCF-7 (ER+/PR+) SK-BR-3 (HER2+) MDA-MB-231 (TNBC)

22. SK-BR-3 cells exhibit highest β1 integrin
Hoechst β1 integrin
3-D2-D
MCF-7 (ER+/PR+) SK-BR-3 (HER2+) MDA-MB-231 (TNBC)

23. Conclusions
• A 3D model for growing cancer cells was
determined and tested
• Colony area, diameter, and circularity change
with time
• Various proteins and integrins are expressed
differently in 2D and 3D culture
• By creating an improved in vitro model, cancer
screening can be improved based on
increased knowledge about cancer phenotype

24. Acknowledgements
Lipke Lab Group
Dr. Elizabeth Lipke
Alexander Hodge
Petra Kerscher
Jennifer Kaczmarek
Dr. Wen Seeto
Yuan Tian
Iman Hassani
Maria Kuchla
Shantanu Pradhan
Sara Head
Nidhi Goel
Funding
AU-CMB
AURIC