Tumor Spheroid Formation
Tumor Spheroid Formation - Related Videos
06:26
PTM Based Tumor Microenvironment Monitoring: A Method for Tracking Changes in ECM Rigidity Using Tumor Spheroids
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2023
This protocol describes a technique for monitoring extracellular matrix rigidity changes by using particle-tracking microrheology (PTM). This is done by culturing 3D pancreatic tumor spheroids into ECM with embedded fluorescent probes for video particle-tracking and analysis.
03:20
Splitting Spheroids for Sub-culturing and Shipping: A Procedure for Propagating and Transferring Spheroids from Small Bowel Neuroendocrine Tumor
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2023
This video demonstrates a strategy to split and propagate spheroids developed from small bowel neuroendocrine tumor cells. These spheroids have small bowel neuroendocrine tumor markers and can be propagated and further used to test anti-cancer drugs.
03:18
Generation of a Three-Dimensional Spheroid Model of Tumor Cell Invasion
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2025
Source: Guyon, J., et al. A 3D Spheroid Model for Glioblastoma. J. Vis. Exp. (2020)The video demonstrates the generation of a three-dimensional spheroid model of tumor cell invasion. Spheroids are formed from human brain tumor cells and embedded in a collagen matrix. The tumor cells at the spheroid periphery adopt an invasive phenotype, secreting proteases to degrade the collagen and advancing to invade the surrounding matrix.
02:02
A Three-Dimensional Co-culture Model of Infiltrating T Cells in Tumor Spheroids
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2025
This video demonstrates a method to establish a three-dimensional tumor spheroid and T cell co-culture. An agarose cast with microwells containing tumor spheroids is prepared, followed by the addition of T cells. After incubation, this setup allows studying the interaction of T cells with tumor spheroids in a controlled three-dimensional environment.
02:38
Characterization of Small Bowel Neuroendocrine Tumor Spheroids Using Immunofluorescence
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2025
Source: Ear, P. H., et. al. Establishment and Characterization of Small Bowel Neuroendocrine Tumor Spheroids. J. Vis. Exp. (2019)This video demonstrates the immunofluorescence staining of small bowel neuroendocrine tumor spheroids. The spheroids are fixed, permeabilized, and blocked and are then incubated with primary and secondary antibodies. A dye is added to stain nuclei before mounting and visualizing them under a fluorescent microscope.
FAQs
Nonadherent or low-attachment conditions reduce the tendency of cancer cells to spread across a culture surface, allowing them to remain together. This supports cell-cell adhesion and the progressive assembly of a multicellular aggregate. In practice, the culture condition is central to obtaining a three-dimensional structure rather than a dispersed or surface-attached population for cancer studies.
Within a tumor spheroid, nutrients, oxygen, and waste are not distributed uniformly. Their movement from the spheroid surface toward its center creates internal gradients, so cells in different locations experience different local conditions. These spatial differences help researchers examine how tumor organization and microenvironmental variation influence cancer-cell behavior and treatment responses.
Compared with conventional two-dimensional culture, Tumor Spheroid Formation preserves three-dimensional cell organization and creates spatial gradients that flat cultures do not represent in the same way. This added organization can provide a more physiologically relevant setting for evaluating growth, invasion, and anticancer treatment responses, while complementing rather than replacing simpler culture systems.
Cell-cell adhesion and proliferation contribute different but connected functions during assembly. Adhesion keeps neighboring cancer cells associated, while proliferation increases the multicellular population within the developing structure. Together, these processes support spheroid growth and organization, allowing investigators to relate aggregate architecture to cancer-cell behavior rather than studying isolated cells alone.
To establish a model, researchers place cancer cells under nonadherent or low-attachment conditions and allow them to remain associated. The cells assemble through cell-cell adhesion and proliferate into a three-dimensional aggregate. The resulting spheroid can then serve as the experimental system for examining growth, invasion, microenvironment effects, or responses to anticancer treatments.
They are useful when the study needs a three-dimensional cancer model that captures organization and internal gradients alongside treatment exposure. Researchers can compare how anticancer treatments affect cells within this structured context, generating information about therapeutic responses that may be more physiologically relevant than observations from conventional two-dimensional cultures alone.
The model supports investigation of cancer-cell growth, invasion, and responses to anticancer treatments, while also enabling study of tumor microenvironment effects. These readouts connect the spheroid’s three-dimensional organization and internal gradients with experimental outcomes, helping researchers assess how spatially structured conditions may shape therapeutic performance and tumor-related behavior.