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Related Experiment Video

Updated: Jul 30, 2025

Assessing Cell Viability and Death in 3D Spheroid Cultures of Cancer Cells
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Cell Viability Assay with 3D Prostate Tumor Spheroids.

Ezgi Oner1,2,3, Steven G Gray2,3, Stephen P Finn4,5,6,7,8

  • 1Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Izmir Katip Celebi University, Balatcik, Izmir, Turkey.

Methods in Molecular Biology (Clifton, N.J.)
|May 18, 2023
PubMed
Summary

This study presents a simplified method for assessing prostate cancer cell viability using 3D tumor spheroids and the WST-8 assay. The protocol streamlines spheroid formation and drug testing for improved 3D in vitro cancer research.

Keywords:
3D in vitro modelsCytotoxicityPolyHEMA methodProstaspheresProstate cancerThree-dimensional cell cultureTumor spheroidsUltra-low attachment surfaceWST-8 (Cell Counting Kit-8, CCK-8) cell viability assay

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Area of Science:

  • Biomedical Sciences
  • Cancer Research
  • Cell Biology

Background:

  • 3D in vitro models offer a more physiologically relevant platform for cancer research compared to 2D cultures.
  • Accurate assessment of cell viability in 3D models is crucial for drug discovery and efficacy testing.
  • Existing methods for 3D spheroid analysis can be complex and labor-intensive.

Purpose of the Study:

  • To describe a streamlined protocol for generating 3D prostate tumor spheroids using the polyHEMA technique.
  • To detail the application of the WST-8 (Cell Counting Kit 8) assay for measuring cell viability in these 3D spheroids.
  • To highlight the advantages of this protocol, including simplified spheroid formation and handling.

Main Methods:

  • Formation of 3D prostate tumor spheroids utilizing the polyHEMA technique.
  • Application of drug treatments directly to the formed spheroids.
  • Quantification of cell viability using the WST-8 assay (also known as Cell Counting Kit 8).

Main Results:

  • Successful generation of 3D prostate tumor spheroids without the need for extracellular matrix components.
  • Demonstration of effective drug treatment and subsequent WST-8 assay application on the spheroids.
  • Calculation of percentage cell viability for PC-3 prostate tumor spheroids.

Conclusions:

  • The described protocol provides an efficient and validated method for assessing cell viability in 3D prostate tumor spheroids.
  • The polyHEMA technique simplifies spheroid formation and eliminates complex transfer steps, making it suitable for various cancer cell lines.
  • This approach enhances the utility of 3D in vitro models for cancer drug screening and research.