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In Vitro 3D Models of Tunable Stiffness
Elysse C Filipe1,2, Amelia L Parker1,2, Antonia L Cadell1
1The Garvan Institute of Medical Research and The Kinghorn Cancer Centre, Darlinghurst, NSW, Australia.
Methods in Molecular Biology (Clifton, N.J.)
|March 20, 2021
Summary
This study introduces a new 3D hydrogel model for cancer research, enabling high-throughput in vitro screening of treatments. The system allows researchers to test drug efficacy on malignant cells within a tunable mechanical environment, mimicking in vivo conditions.
Area of Science:
- Biomedical Engineering
- Cancer Biology
- Materials Science
Background:
- Three-dimensional spheroid models are crucial for assessing cancer cell colony formation in vitro.
- These models serve as surrogates for in vivo testing, facilitating large-scale treatment condition analysis.
- The tumor microenvironment's mechanical properties, altered by extracellular matrix changes, significantly impact cancer progression.
Purpose of the Study:
- To present an adapted, high-throughput protocol for spheroid formation using a composite alginate hydrogel system.
- To establish a biomechanically tunable 3D environment for studying cancer cell behavior.
- To enable examination of treatment effects on cells within hydrogels of defined stiffness, mimicking the cancer microenvironment.
Main Methods:
- Development of a high-throughput method for creating homogeneous interpenetrating polymer networks of collagen and alginate.
- Utilizing a composite alginate hydrogel system for spheroid formation in a tunable 3D environment.
- Embedding malignant cell lines (breast cancer, lung cancer, melanoma) within the hydrogel for spheroid culture.
Main Results:
- The composite hydrogel system successfully supports single-cell spheroid formation in multiple cancer cell lines.
- Spheroids formed within the hydrogel can be robustly analyzed for key colony formation metrics.
- Analyzed metrics include spheroid size, spheroid number, and overall cell viability.
Conclusions:
- This protocol offers a high-throughput, accessible method for in vitro cancer screening.
- The tunable 3D hydrogel environment allows for controlled biomechanical background analysis.
- The system facilitates robust interrogation of treatment efficacy against cancer spheroids.
