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Updated: Mar 6, 2026
Generation of High-Throughput Three-Dimensional Tumor Spheroids for Drug Screening
Published on: September 5, 2018
3D Models of the NCI60 Cell Lines for Screening Oncology Compounds
Mike Selby1, Rene Delosh1, Julie Laudeman1
11 Applied/Developmental Research Directorate, Leidos Biomedical Research, Inc., Frederick National Lab for Cancer Research, Frederick, MD, USA.
Abstract:
The NCI60 cell line panel screen includes 60 human tumor cell lines derived from nine tumor types that has been used over the past 20+ years to screen small molecules, biologics, and natural products for activity. Cells in monolayer culture in 96-well plates are exposed to compounds for 48 h, and Sulforhodamine B is used to determine cell viability. Data analysis tools such as COMPARE allow classification of compounds based on the pattern of cell line response. However, many compounds highly active in monolayer cell culture fail to show efficacy in vivo. Therefore, we explored 3D culture of the NCI60 panel as a strategy to improve the predictive accuracy of the screen. 3D cultures more closely resemble tumors than monolayer cultures with tighter cell-cell contact and nutrient and oxygen gradients between the periphery and the center. We optimized the NCI60 cell line panel for generating 3D spheroids of a prespecified diameter (300-500 µm) in ultra-low attachment (ULA) plates. Spheroids were classified into four categories based on imaging, and concentration response of select agents in 2D and 3D models is presented.
Insights
The NCI60 cell line panel now uses 3D spheroid cultures to better predict drug efficacy. This advanced screening method improves upon traditional 2D monolayer cultures for cancer drug discovery.
Area of Science:
- Oncology
- Drug Discovery
- Cell Biology
Background:
- The NCI60 cell line panel is a widely used resource for screening anticancer compounds.
- Traditional 2D monolayer cell cultures have limitations in predicting in vivo drug efficacy.
- Many promising compounds fail in clinical trials despite high activity in 2D screens.
Purpose of the Study:
- To enhance the predictive accuracy of the NCI60 cell line panel screen.
- To explore the utility of 3D spheroid cultures as a more physiologically relevant model.
- To optimize 3D culture conditions for the NCI60 panel.
Main Methods:
- Optimization of NCI60 cell line panel for 3D spheroid generation (300-500 µm diameter).
- Utilization of ultra-low attachment (ULA) plates for spheroid formation.
- Assessment of cell viability using Sulforhodamine B in both 2D and 3D cultures.
- Classification of spheroids into four categories based on imaging analysis.
Main Results:
- Established reproducible 3D spheroid formation for the NCI60 panel.
- Demonstrated differences in compound response between 2D and 3D NCI60 models.
- Presented concentration-response data for select agents in both culture formats.
Conclusions:
- 3D spheroid culture of the NCI60 panel offers a more predictive model for anticancer drug screening.
- This approach more closely mimics the tumor microenvironment compared to 2D cultures.
- The optimized 3D NCI60 system has the potential to improve the success rate of drug discovery.
