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Updated: Jul 20, 2026
Generation of High-Throughput Three-Dimensional Tumor Spheroids for Drug Screening
Published on: September 5, 2018
In vitro throughput screening of anticancer drugs using patient-derived cell lines cultured on vascularized
Yuki Takahashi1, Rii Morimura1, Kei Tsukamoto2
1Business Development Division, Technical Research Institute, TOPPAN Holdings Inc., Saitama 345-8508, Japan; Division of Clinical Chemotherapy, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, Tokyo 135-8550, Japan.
Abstract:
The development of high-throughput anticancer drug screening methods using patient-derived cancer cell (PDC) lines that maintain their original characteristics in an in vitro three-dimensional (3D) culture system poses a significant challenge to achieving personalized cancer medicine. Because stromal tissue plays a critical role in the composition and maintenance of the cancer microenvironment, in vitro 3D-culture using reconstructed stromal tissues has attracted considerable attention. Here, a simple and unique in vitro 3D-culture method using heparin and collagen together with fibroblasts and endothelial cells to fabricate vascularized 3D-stromal tissues for in vitro culture of PDCs is reported. Whereas co-treatment with bevacizumab, a monoclonal antibody against vascular endothelial growth factor, and 5-fluorouracil significantly reduced the survival rate of 3D-cultured PDCs to 30%, separate addition of each drug did not induce comparable strong cytotoxicity, suggesting the possibility of evaluating the combined effect of anticancer drugs and angiogenesis inhibitors. Surprisingly, drug evaluation using eight PDC lines with the 3D-culture method resulted in a drug efficacy concordance rate of 75% with clinical outcomes. The model is expected to be applicable to in vitro throughput drug screening for the development of personalized cancer medicine. STATEMENT OF SIGNIFICANCE: To replicate the cancer microenvironment, we constructed a cancer-stromal tissue model in which cancer cells are placed above and inside stromal tissue with vascular network structures derived from vascular endothelial cells in fibroblast tissue using CAViTs method. Using this method, we were able to reproduce the invasion and metastasis processes of cancer cells observed in vivo. Using patient-derived cancer cells, we assessed the possibility of evaluating the combined effect with an angiogenesis inhibitor. Further, primary cancer cells also grew on the stromal tissues with the normal medium. These data suggest that the model may be useful for new in vitro drug screening and personalized cancer medicine.
Insights
This study introduces a novel 3D culture model using patient-derived cancer cells and vascularized stromal tissue. This innovative method accurately predicts anticancer drug efficacy, advancing personalized cancer medicine.
Area of Science:
- Biomedical Engineering
- Cancer Biology
- Drug Discovery
Background:
- Personalized cancer medicine requires in vitro models that mimic the patient's tumor microenvironment.
- Existing 3D culture methods face challenges in replicating the complex interactions within the tumor microenvironment.
Purpose of the Study:
- To develop a novel in vitro 3D culture system for patient-derived cancer cells (PDCs).
- To create vascularized 3D-stromal tissues that replicate the cancer microenvironment.
- To evaluate the efficacy of anticancer drugs and angiogenesis inhibitors in a 3D culture system.
Main Methods:
- Fabrication of vascularized 3D-stromal tissues using heparin, collagen, fibroblasts, and endothelial cells.
- Culture of PDCs within the fabricated 3D-stromal tissues.
- Co-treatment of 3D-cultured PDCs with bevacizumab and 5-fluorouracil.
- Evaluation of drug efficacy concordance with clinical outcomes for eight PDC lines.
Main Results:
- The 3D culture system successfully replicated cancer cell invasion and metastasis.
- Combined treatment with bevacizumab and 5-fluorouracil significantly reduced PDC survival.
- The 3D-culture method demonstrated a 75% concordance rate with clinical outcomes for drug efficacy.
- Primary cancer cells grew effectively on the stromal tissues in normal medium.
Conclusions:
- The developed 3D vascularized stromal tissue model is a promising tool for in vitro drug screening.
- This model facilitates the evaluation of combined drug effects, including angiogenesis inhibitors.
- The system holds significant potential for advancing personalized cancer medicine through accurate drug response prediction.
