kelsey henderson cmb usrs presentation
TRANSCRIPT
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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
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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
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Breast Cancer Molecular Subtypes
http://labmed.ascpjournals.org/content/41/6/364.figures-only
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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)
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3D Cell Culture: Cells Encapsulated in Hydrogels
http://pubs.rsc.org/en/content/articlehtml/2013/cs/c3cs60040h
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MCF-7 (ER+/PR+)
SK-BR-3 (HER2+)
MDA-MB-231 (TNBC)
Breast cancer cells encapsulated in hydrogels on day 6
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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
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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
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MCF-7 progression of encapsulated cells
Day 0 Day 3 Day 15
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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)
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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
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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+)
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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
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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+)
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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
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Cell structure and morphologyHoechst Phalloidin
MCF-7 (ER+/PR+) MDA-MB-231 (TNBC)SK-BR-3 (HER2+)
2-D
3-D
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Cell proliferationHoechst Ki67
2-D
3-D
MCF-7 (ER+/PR+) SK-BR-3 (HER2+) MDA-MB-231 (TNBC)
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Epithelial vs. Mesenchymal Cells
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Mesenchymal cells express higher vimentinHoechst Vimentin
3-D
2-D
MCF-7 (ER+/PR+) SK-BR-3 (HER2+) MDA-MB-231 (TNBC)
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Epithelial cells express more E-CadherinHoechst E-Cadherin
3-D
MCF-7 (ER+/PR+) SK-BR-3 (HER2+) MDA-MB-231 (TNBC)
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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)
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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)
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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
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AcknowledgementsLipke Lab GroupDr. Elizabeth LipkeAlexander HodgePetra KerscherJennifer KaczmarekDr. Wen SeetoYuan TianIman HassaniMaria KuchlaShantanu PradhanSara HeadNidhi Goel
FundingAU-CMBAURIC