Description
- Abstract:
- The epithelial-mesenchymal transition (EMT) is associated with solid tumor dissemination and metastasis. Directed migration of tumor cells into the surrounding stroma occurs through both collective and individual phenotypes, mediated by varying cell-cell and cell-matrix adhesions. Such invasive phenotypes may utilize spatially and temporally coordinated tractions to deform and remodel extracellular matrix. However, the mechanistic role of EMT in collective and individual invasion remains poorly understood. We utilized the hanging drop method to aggregate mammary epithelial (MCF-10A) cells, with an inducible gene expression construct that induces EMT master regulator Snail-1 in response to stimulation of the estrogen receptor with tamoxifen. Spheroids were transferred into 3D matrix (e.g., bovine collagen I) embedded with tracer particles to create a spheroid invasion model. Over several days, we observed and imaged spheroid invasion profiles using spinning disk confocal microscopy with environmental control, and 3D single cell and particle tracking was performed using custom MATLAB code. Our findings revealed that induction of EMT in MCF-10A spheroids via Snail-1 upregulation decreased cell-cell adhesions, resulting in the loss of leader-follower cell behavior and collective invasion. EMT induction was also associated with a loss of intra-spheroid coordinated cell motion as well as spheroid generated matrix deformations. The use of confocal imaging and 3D cell culture techniques allowed for high-resolution characterization of collective and individual cell dynamics. These findings have intriguing implications for cancer research and therapy development, including EMT as a potential therapeutic target.
- Notes:
- Thesis (Sc. M.)--Brown University, 2023
Citation
Marzoratti, Alejandro M.,
"Collective and Individual Invasion of Multicellular Spheroids in 3D Matrix"
(2023).
Biomedical Engineering Theses and Dissertations.
Brown Digital Repository. Brown University Library.
https://repository.library.brown.edu/studio/item/bdr:z7n9xtj6/
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Biomedical Engineering Theses and Dissertations
Theses and Dissertations for the Biomedical Engineering department....