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Updated: Dec 24, 2025
Generation of 3D Tumor Spheroids for Drug Evaluation Studies
Published on: February 24, 2023
Selective Cytotoxicity of a Novel Trp-Rich Peptide against Lung Tumor Spheroids Encapsulated inside a 3D Microfluidic
Nandini Dhiman1,2, Nadin Shagaghi1, Mrinal Bhave1
1Department of Chemistry and Biotechnology, Faculty of Science, Engineering and Technology, Swinburne University of Technology, Hawthorn, Victoria, 3122, Australia.
Abstract:
There is a globally rising healthcare need to develop new anticancer therapies as well as to test them on biologically relevant in vitro cancer models instead of overly simplistic 2D models. To address both these needs, a 3D lung cancer spheroid model is developed using human A549 cells trapped inside a collagen gel in a compartmentalized microfluidic device and homogenously sized (35-45 µm) multicellular tumor spheroids are obtained in 5 days. The novel tryptophan-rich peptide P1, identified earlier as a potential anticancer peptide (ACP), shows enhanced cytotoxic efficacy against A549 tumor spheroids (>75%) in clinically relevant low concentrations, while it does not affect human amniotic membrane mesenchymal stem cells at the same concentrations (<15%). The peptide also inhibits the formation of tumor spheroids by reducing cell viability as well as lowering the proliferative capacity, which is confirmed by the expression of cell proliferation marker Ki-67. The ACP offers a novel therapeutic strategy against lung cancer cells without affecting healthy cells. The microfluidic device used is likely to be useful in helping develop models for several other cancer types to test new anticancer agents.
Insights
A novel tryptophan-rich peptide P1 demonstrates significant anticancer efficacy against lung cancer spheroids in a 3D microfluidic model. This peptide selectively targets cancer cells, offering a promising new therapeutic strategy without harming healthy stem cells.
Area of Science:
- Oncology
- Biotechnology
- Biomaterials
Background:
- Rising global need for effective anticancer therapies and biologically relevant in vitro cancer models.
- Limitations of traditional 2D cell culture models for cancer research.
- Development of 3D multicellular tumor spheroids as advanced cancer models.
Purpose of the Study:
- To develop a 3D lung cancer spheroid model using a microfluidic device.
- To evaluate the anticancer efficacy of a novel tryptophan-rich peptide P1.
- To assess the peptide's safety on healthy human cells.
Main Methods:
- Creation of a 3D lung cancer spheroid model using human A549 cells in a collagen gel within a microfluidic device.
- Treatment of spheroids with peptide P1 at low concentrations.
- Assessment of cell viability, proliferation (Ki-67 expression), and cytotoxicity.
- Evaluation of peptide P1's effect on human amniotic membrane mesenchymal stem cells.
Main Results:
- Homogeneously sized (35-45 µm) multicellular tumor spheroids were obtained in 5 days.
- Peptide P1 showed >75% cytotoxic efficacy against A549 tumor spheroids at low concentrations.
- Peptide P1 exhibited <15% cytotoxicity against human amniotic membrane mesenchymal stem cells.
- Peptide P1 inhibited spheroid formation, reduced cell viability, and lowered proliferative capacity.
Conclusions:
- Peptide P1 is a potent anticancer peptide (ACP) with selective efficacy against lung cancer cells.
- The developed 3D microfluidic model provides a biologically relevant platform for testing anticancer agents.
- Peptide P1 represents a novel therapeutic strategy for lung cancer with minimal impact on healthy cells.
