3d Spheroid Formation

3D spheroid formation is a bioengineering method for organizing living cells into compact, three-dimensional aggregates that better reproduce aspects of tissue structure than conventional two-dimensional cultures. Cells are typically placed under nonadherent conditions, such as low-attachment surfaces or hanging drops, where cell-cell adhesion and extracellular matrix interactions drive aggregation, compaction, and self-assembly into spheroids. These constructs develop gradients of oxygen, nutrients, and metabolites, creating microenvironments that can influence proliferation, differentiation, and drug responses. Spheroids support tissue engineering, cancer research, regenerative medicine, and toxicity testing, while providing a scalable platform for studying cell behavior and evaluating biomaterials or therapeutic strategies.

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Research

JoVE Journal - Bioengineering

Creation of Cardiac Tissue Exhibiting Mechanical Integration of Spheroids Using 3D Bioprinting

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Cited by 55 •

2017

This protocol describes 3D bioprinting of cardiac tissue without the use of biomaterials. 3D bioprinted cardiac patches exhibit mechanical integration of component spheroids and are highly promising in cardiac tissue regeneration and as 3D models of heart disease.

Extracellular Flux Assay in 3D Spheroids: A Method for Measuring the Metabolic Activity of Pancreatic Tumor Spheroids

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2023

This protocol describes the technique for performing extracellular flux assay on 3D spheroids generated from the co-culture of pancreatic cancer cells (Patu8902) and activated pancreatic fibroblasts (PS1) to measure the two major energy-producing pathways, glycolysis and mitochondrial respiration.

Spheroid Generation from Cell Lines: A Three-Dimensional (3D) Cell Culture Method

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2023

Spheroids are a type of 3D cell culture model used to more accurately represent a cell’s environment compared to 2D cell culture monolayer models. In the example protocol, we will see scientists generate spheroids from a colorectal cancer cell line and apply live-cell imaging to track the spheroids’ growth.

Evaporation-reducing Culture Condition Increases the Reproducibility of Multicellular Spheroid Formation in Microtiter Plates

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Cited by 6 •

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.

An Efficient and Flexible Cell Aggregation Method for 3D Spheroid Production

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Cited by 67 •

2017

Here, we describe a rapid and flexible protocol for the formation of 3D cell spheroids through cell aggregation. This is easily adapted to multiple cell types and is suitable for use in a variety of applications including cell migration, invasion, or anoikis assays, and for imaging and quantifying cell-matrix interactions.

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