Spheroid Growth Assays
In vitro culture models of cancer cells are integral to pre-clinical drug development and advancing our understanding of cancer cell biology. While still relatively new compared to traditional 2D monolayer cultures, 3D cell culture models have emerged as advanced tools to accelerate drug discovery efforts.
However, many available three-dimensional techniques for generating and quantitatively analyzing spheroids are time consuming, laborious, costly, and/or lack reproducibility, prohibiting the effective utilization of these more stringent and representative in vitro tumor models.
It is well established that conventional monolayer cultures of tumor cells on plastic do not adequately reflect the in vivo situation: Cancer cells within a solid tumor are part of a complex 3-dimensional (3D) structure with a specific organization and cytoarchitecture. Tumor cells interact with each other and with the surrounding microenvironment. These complex interactions in combination with the unique properties of a 3D structure affect tumor cell properties and behavior, gene expression, distribution of proteins, and ultimately, response to anticancer drugs. Thus, 2D culture models have limited value for investigating tumor pathophysiologic pathways or predicting the potential efficacy of drug candidates.
To develop tumor models that better mimic the complexity and heterogeneity of clinical tumors, 3D culture methods have been developed in which tumor cells are grown as multicell aggregates, or spheroids. There are a multitude of approaches to generating these models, and can generally be described as liquid-based or scaffold-based and detailed elsewhere1. Liquid-based models are easily achieved using round-bottom ultra-low attachment (ULA) microplates to promote spheroid self-assembly. These models, generally yielding a single tumor spheroid per well, exhibit key features of solid tumors, with larger spheroids consisting of proliferating, quiescent, and necrotic zones resulting from a radial gradient of nutrients, metabolites and oxygen2. Scaffold-based models used today primarily rely on an extracellular matrix (ECM) such as Matrigel or collagen to recapitulate both physical and biochemical characteristic of the tumor microenvironment. Generally, these models are generated using flat bottom plates and result in multiple tumor spheroids per well.
Effective analysis of 3D tumor spheroids can be challenging. These heterogeneous models are multi-faceted and dynamic. Traditional plate reader assays are endpoint and lack multiple aspects of image-based analysis, including morphological information and ability to confirm data within images. Conventional imaging systems are inherently difficult to adapt to kinetic analyses of in vitro culture models, suffering from:
Incucyte® 3D Tumor Spheroid Assays offer an integrated turnkey solution to automatically monitor and quantify tumor spheroid formation, growth and health in real time inside your tissue culture incubator.
An integrated solution to automatically track and quantify tumor spheroid formation, growth and death in real time and in 96- and 384-well formats — inside your tissue culture incubator.
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An integrated turnkey solution to automatically track and quantify tumor spheroid formation, growth and health in real time inside your tissue culture incubator.
Development and Optimization of Matrigel-Based Multi-Spheroid 3d Tumor Assays Using Real-Time Live-Cell Analysis
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