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Updated: Aug 1, 2025
Therapy Testing in a Spheroid-based 3D Cell Culture Model for Head and Neck Squamous Cell Carcinoma
Published on: April 20, 2018
3D spheroid culture to examine adaptive therapy response in invading tumor cells
Felix Weiss1, Nader Atlasy2, Vince van Reijmersdal1
1Department of Cell Biology, Radboud University Medical Centre, P.O. Box 9101, 6525 GA Nijmegen, The Netherlands.
Abstract:
3D in vitro culture models of cancer cells in extracellular matrix (ECM) have been developed to investigate drug targeting and resistance or, alternatively, mechanisms of invasion; however, models allowing analysis of shared pathways mediating invasion and therapy resistance are lacking. To evaluate therapy response associated with cancer cell invasion, we here used 3D invasion culture of tumor spheroids in 3D fibrillar collagen and applied Ethanol-Ethyl cinnamate (EtOH-ECi) based optical clearing to detect both spheroid core and invasion zone by subcellular-resolved 3D microscopy. When subjected to a single dose of irradiation (4 Gy), we detected significant cell survival in the invasion zone. By physical separation of the core and invasion zone, we identified differentially regulated genes preferentially engaged in invading cells controlling cell division, repair, and survival. This imaging-based 3D invasion culture may be useful for the analysis of complex therapy-response patterns in cancer cells in drug discovery and invasion-associated resistance development.
Supplementary Information:
The online version contains supplementary material available at 10.1007/s44164-022-00040-x.
Insights
A new 3D cancer model reveals that invading cancer cells survive radiation therapy better than core cells. This discovery aids in understanding invasion-associated therapy resistance and drug discovery.
Area of Science:
- Oncology
- Biotechnology
- Cell Biology
Background:
- 3D in vitro cancer models using extracellular matrix (ECM) are crucial for studying drug resistance and invasion.
- Existing models often fail to analyze shared pathways linking cancer cell invasion and therapy resistance simultaneously.
Purpose of the Study:
- To develop and utilize a novel 3D invasion culture model for evaluating therapy response in relation to cancer cell invasion.
- To investigate the mechanisms underlying differential therapy response between spheroid core and invasive cancer cells.
Main Methods:
- Established a 3D invasion culture model using tumor spheroids in 3D fibrillar collagen.
- Applied Ethanol-Ethyl cinnamate (EtOH-ECi) based optical clearing for subcellular-resolved 3D microscopy.
- Physically separated spheroid core and invasion zones for gene expression analysis.
Main Results:
- Observed significant cell survival in the invasion zone after a single dose of 4 Gy irradiation.
- Identified differentially regulated genes in invading cells, particularly those involved in cell division, repair, and survival.
- Demonstrated the utility of the 3D model in analyzing complex therapy response patterns.
Conclusions:
- The developed 3D invasion culture model effectively visualizes and analyzes therapy response in both spheroid cores and invasive zones.
- Invading cancer cells exhibit enhanced survival mechanisms, contributing to invasion-associated therapy resistance.
- This model holds potential for advancing drug discovery and understanding resistance mechanisms in cancer research.
