Spheroid Formation
Spheroid Formation - Related Videos
11:24
Evaporation-reducing Culture Condition Increases the Reproducibility of Multicellular Spheroid Formation in Microtiter Plates
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Cited by 9 •
2017
The uneven loss of medium from microtiter plates affects the reproducibility of uniform multicellular tumor spheroid formation. Improving culture conditions to reduce significant medium loss will improve the reproducibility of spheroid formation and the results of spheroid-based assays using the liquid-overlay technique.
10:44
Tumor Spheroid Fabrication and Encapsulation in Polyethylene Glycol Hydrogels for Studying Spheroid-Matrix Interactions
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Cited by 1 •
2023
Here, we present a protocol that enables fast, robust, and cheap fabrication of tumor spheroids followed by hydrogel encapsulation. It is widely applicable as it does not require specialized equipment. It would be particularly useful for exploring spheroid-matrix interactions and building in vitro tissue physiology or pathology models.
02:42
Preparation of Cancer Spheroid Blocks by Flash Freezing: A Procedure To Preserve Cancer Spheroids in Frozen Matrix Blocks
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2023
This video demonstrates the preparation of cancer spheroid blocks by flash freezing, a technique in which a tissue sample of interest is frozen to an ultracold temperature, restraining the spheroids inside the matrix.
12:11
Using Optical Tweezers for the Generation of Hybrid Spheroids
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2025
In this paper, we propose a protocol for hybrid spheroid manufacturing using optical tweezers. This method allows precise control of the early stages of the spheroid formation process, which is unattainable by other methods.
08:27
Mechanical Mapping of Spheroids Using Brillouin Spectroscopy
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2025
This article describes a protocol for mechanical characterisation of living spheroids within a 3D matrix using Brillouin micro-spectroscopy. This all-optical method enables spheroid visualisation with microscale resolution and quantitative mechanical properties. This approach has implications for mechanical phenotyping; for instance, determining pathological states of tumor spheroids within a 3D microenvironment.
FAQs
Cell-cell adhesion drives the transition from dispersed cells to compact spheroids. Under nonadherent or low-attachment conditions, cells preferentially interact with one another rather than spreading across a surface, allowing aggregates to tighten into spherical structures. This organization creates internal gradients of nutrients, oxygen, and signaling molecules, which can produce spatially different cellular conditions within the same construct.
Spheroid formation preserves cell-cell interactions and three-dimensional organization that flat cultures cannot reproduce as effectively. The resulting architecture offers a more tissue-like setting for examining how cells respond to neighboring cells, changing oxygen or nutrient availability, and local signals. In bioengineering, this distinction helps researchers choose a model when two-dimensional growth may not reflect native tissue behavior.
Internal gradients make a spheroid biologically nonuniform rather than equivalent throughout. Differences in oxygen, nutrients, and signaling molecules can shape local cell behavior and may affect viability, differentiation, and responses to drugs or biomaterials. Interpreting results therefore requires considering where cells are located within the aggregate, not only the overall spheroid response.
To generate spheroids, cells are placed under nonadherent or low-attachment conditions and allowed to assemble through cell-cell adhesion. As clustering proceeds, aggregates compact and become spherical. Researchers can then investigate the resulting structures by examining cell interactions, organization, viability, differentiation, or responses to selected drugs and biomaterials, depending on the experimental question.
Researchers use spheroids when a three-dimensional model is needed to study tissue organization or cell behavior more realistically than a flat culture. They can apply the model to investigate disease progression, test responses to drugs or biomaterials, and examine how cells interact within an engineered tissue context. The appropriate outcome depends on the biological or bioengineering question.
Within bioengineering, spheroids support tissue engineering and regenerative medicine by providing a controlled way to study cell interactions and tissue organization. They also help researchers examine viability, differentiation, and the development of more physiologically relevant engineered tissues. This makes the approach useful when the goal is to connect cellular behavior with how engineered constructs may organize and function.