Embed Size (px)
Citation preview
Development of 3D Breast Cancer Models in PEG-Fibrinogen Hydrogels Kelsey Henderson, Shantanu Pradhan, Dr. Elizabeth A Lipke
Department of Chemical EngineeringAuburn University, Auburn, AL, USA
July 24, 2015
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
Breast Cancer Molecular Subtypes
http://labmed.ascpjournals.org/content/41/6/364.figures-only
2D Cell Culture
Problem: Not accurate depiction of how cancer cells grow in the body
https://www.spectrumchemical.com/OA_HTML/lab-supplies-products_CorningR-Sterile-Cell-Culture-Flasks_305276.jsp?section=23372
MCF-7 (ER+/PR+) SK-BR-3 (HER2+) MDA-MB-231 (TNBC)
3D Cell Culture: Cells Encapsulated in Hydrogels
http://pubs.rsc.org/en/content/articlehtml/2013/cs/c3cs60040h
MCF-7 (ER+/PR+)
SK-BR-3 (HER2+)
MDA-MB-231 (TNBC)
Breast cancer cells encapsulated in hydrogels on day 6
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
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
MCF-7 progression of encapsulated cells
Day 0 Day 3 Day 15
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)
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
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+)
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
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+)
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
Cell structure and morphologyHoechst Phalloidin
MCF-7 (ER+/PR+) MDA-MB-231 (TNBC)SK-BR-3 (HER2+)
2-D
3-D
Cell proliferationHoechst Ki67
2-D
3-D
MCF-7 (ER+/PR+) SK-BR-3 (HER2+) MDA-MB-231 (TNBC)
Epithelial vs. Mesenchymal Cells
Mesenchymal cells express higher vimentinHoechst Vimentin
3-D
2-D
MCF-7 (ER+/PR+) SK-BR-3 (HER2+) MDA-MB-231 (TNBC)
Epithelial cells express more E-CadherinHoechst E-Cadherin
3-D
MCF-7 (ER+/PR+) SK-BR-3 (HER2+) MDA-MB-231 (TNBC)
MDA-MB-231 cells exhibit highest αvβ3 Hoechst αvβ3
3-D
2-D
MCF-7 (ER+/PR+) SK-BR-3 (HER2+) MDA-MB-231 (TNBC)
SK-BR-3 cells exhibit highest β1 integrinHoechst β1 integrin
3-D
2-D
MCF-7 (ER+/PR+) SK-BR-3 (HER2+) MDA-MB-231 (TNBC)
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
AcknowledgementsLipke Lab GroupDr. Elizabeth LipkeAlexander HodgePetra KerscherJennifer KaczmarekDr. Wen SeetoYuan TianIman HassaniMaria KuchlaShantanu PradhanSara HeadNidhi Goel
FundingAU-CMBAURIC
