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Updated: Jul 31, 2025

Assessing Cell Viability and Death in 3D Spheroid Cultures of Cancer Cells

Assessing Cell Viability and Death in 3D Spheroid Cultures of Cancer Cells

Published on: June 16, 2019

Cell Viability Assays for 3D Cellular Constructs.

Zachary Congress¹, Matthew Brovold¹, Shay Soker²

¹Wake Forest Institute for Regenerative Medicine, Winston-Salem, NC, USA.

Methods in Molecular Biology (Clifton, N.J.)

|May 4, 2023

Summary

This summary is machine-generated.

Traditional 2D cell cultures do not accurately mimic in vivo conditions. This chapter reviews cell viability assays essential for evaluating health in advanced 3D tissue constructs for improved drug development.

Keywords:

3D assays ATP Biofabrication Bioprinting Cell culture models Cell viability ECM Hydrogels Organoids Proliferation

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Last Updated: Jul 31, 2025

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Area of Science:

Biomedical Engineering
Cell Biology
Drug Development

Background:

Traditional two-dimensional (2D) cell cultures lack the complex microenvironment of in vivo tissues.
2D cultures inadequately replicate cell-to-cell interactions, extracellular matrix (ECM) components, and topographical/stiffness cues.
These limitations in 2D models can alter cell physiology and molecular properties, impacting research accuracy.

Purpose of the Study:

To highlight the limitations of 2D cell cultures in mimicking in vivo conditions.
To introduce the necessity of advanced 3D tissue constructs for more accurate physiological modeling.
To review essential cell viability assays for assessing cell health in 3D environments.

Main Methods:

Discussion of limitations in 2D cell culture models.
Introduction of biofabrication technologies for 3D tissue construct development.
Overview of cell viability assays adapted for 3D cellular systems.

Main Results:

3D tissue constructs offer a more physiologically relevant environment for cell growth and interaction.
Accurate assessment of cell viability in 3D models is crucial for interpreting experimental outcomes.
Established cell viability assays need adaptation for reliable use with 3D constructs.

Conclusions:

3D cell culture systems represent a significant advancement over 2D models for biomedical research.
Cell viability assays are critical tools for evaluating the efficacy of treatments and the health of 3D tissue constructs.
Standardizing cell viability analysis in 3D environments is essential as these models become the new standard.