1. In Vitro 3D Models of Tunable Stiffness.
- Author
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Filipe EC, Parker AL, Cadell AL, Major G, Croucher DR, and Cox TR
- Subjects
- Alginates chemistry, Animals, Cell Line, Tumor, Collagen chemistry, Drug Screening Assays, Antitumor methods, Extracellular Matrix chemistry, Extracellular Matrix metabolism, Humans, Hydrogels chemistry, Spheroids, Cellular metabolism, Stress, Mechanical, Cell Culture Techniques methods, High-Throughput Screening Assays methods, Spheroids, Cellular cytology, Tissue Scaffolds chemistry
- Abstract
Three-dimensional models of spheroid formation have been routinely used in the cancer field to test the colony forming capacity of malignant cells in an in vitro setting. Use of such a model provides a robust surrogate for in vivo testing, enabling large-scale interrogation into the effect of certain treatment conditions. This adapted protocol describes a high throughput and readily accessible composite alginate hydrogel system for spheroid formation, within a biomechanically tunable three-dimensional environment. This model therefore allows users to examine the effect of certain treatment conditions while cells are embedded within a hydrogel of defined stiffness. This is particularly important in the context of cancer where cells experience a wide range of mechanical properties within their microenvironment, driven by widespread changes in the extracellular matrix composition and architecture.This protocol describes a high-throughput method which results in homogeneous interpenetrating polymer networks of collagen and alginate. We show that this network readily supports single-cell spheroid formation in numerous malignant cell lines (breast cancer, lung cancer, and melanoma) and that these can be robustly analyzed for colony formation measures such as spheroid size, spheroid number, and overall cell viability; therefore, allowing users to undertake high-throughput, in vitro screening against a controlled biomechanical background.
- Published
- 2021
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