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Updated: Jun 10, 2025
Monitoring Cancer Cell Invasion and T-Cell Cytotoxicity in 3D Culture
Published on: June 23, 2020
3D Scaffold-Based Culture System Enhances Preclinical Evaluation of Natural Killer Cell Therapy in A549 Lung Cancer
Eun Hee Han1,2, Sun-Hee Cho1, Sang Nam Lee3
1Biopharmaceutical Research Center, Ochang Institute of Biological and Environmental Science, Korea Basic Science Institute (KBSI), Cheongju 28119, Republic of Korea.
Abstract:
Cell-based immunotherapies have emerged as promising cancer treatment modalities, demonstrating remarkable clinical efficacy. As interest in applying immune cell-based therapies to solid tumors has gained momentum, experimental models that enable long-term monitoring and mimic clinical administration are increasingly necessary. This study explores the potential of scaffold-based cell culture technologies, specifically three-dimensional (3D) extracellular matrix (ECM)-like frameworks, as promising solutions. These frameworks facilitate unhindered immune cell growth and enable continuous cancer cell culture. The three-dimensional (3D) cell culture model was developed using tailored scaffolds for natural killer (NK) cell culture. Within this framework, A549 lung cancer cells were cocultured with NK cells, allowing real-time monitoring for up to 28 days. The expression of critical markers associated with anticancer drug resistance and epithelial-mesenchymal transition (EMT) was evaluated in cancer cells within this 3D culture context. Compared to conventional 2D monolayer cultures, this 3D scaffold-based culture revealed that solid tumor cells, specifically A549 cells, exhibited heightened resistance to anticancer drugs. Additionally, the 3D culture environment upregulated the expression of EMT markers namely vimentin, N-cadherin, and fibronectin, while NK and zEGFR-CAR-NK cells displayed anticancer effects. In the two-dimensional (2D) coculture, only zEGFR-CAR-NK cells exhibited such effects in the 3D coculture system, highlighting an intriguing inconsistency with the 2D culture model, further confirmed by in vivo experiments. This in vitro 3D cell culture model reliably predicts outcomes in NK immunotherapy experiments. Thus, it represents a valuable tool for investigating drug resistance mechanisms and assessing the efficacy of immune cell-based therapies. By bridging the gap between in vitro and in vivo investigations, this model effectively translates potential treatments into animal models and facilitates rigorous preclinical evaluations.
Insights
This study introduces a 3D scaffold model for evaluating cell-based immunotherapies against solid tumors. The advanced 3D culture system accurately predicts drug resistance and immunotherapy outcomes, outperforming traditional 2D models.
Area of Science:
- Biotechnology
- Cancer Research
- Immunotherapy
Background:
- Cell-based immunotherapies show promise for cancer treatment.
- There is a growing need for experimental models that mimic clinical settings for solid tumors.
Purpose of the Study:
- To explore scaffold-based 3D cell culture for long-term monitoring of immune cell therapies.
- To assess drug resistance and epithelial-mesenchymal transition (EMT) in lung cancer cells within a 3D culture system.
Main Methods:
- Developed a 3D scaffold for natural killer (NK) cell culture.
- Cocultured A549 lung cancer cells with NK cells in the 3D scaffold for 28 days.
- Evaluated drug resistance and EMT markers, comparing 3D to 2D cultures.
Main Results:
- 3D cultures showed increased drug resistance in A549 cells compared to 2D cultures.
- EMT markers (vimentin, N-cadherin, fibronectin) were upregulated in 3D.
- NK and zEGFR-CAR-NK cells demonstrated anticancer effects, with specific outcomes differing between 2D and 3D models.
Conclusions:
- The 3D scaffold model reliably predicts NK immunotherapy outcomes, bridging in vitro and in vivo studies.
- This model is valuable for investigating drug resistance and assessing immunotherapy efficacy in preclinical settings.
