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Updated: Mar 22, 2026
Establishing 3-Dimensional Spheroids from Patient-Derived Tumor Samples and Evaluating their Sensitivity to Drugs
Published on: December 16, 2022
Formation of multicellular tumor spheroids induced by cyclic RGD-peptides and use for anticancer drug testing in
Roman Akasov1, Daria Zaytseva-Zotova2, Sergey Burov3
1Polymers for Biology Laboratory, Shemyakin-Ovchinnikov Institute of Bioorganic chemistry of the Russian Academy of Sciences, 117997, Miklukho-Maklaya 16/10, Moscow, Russia; CNRS UMR 7199, University of Strasbourg, Laboratoire de Conception et Application de Molécules Bioactives, 74 route du Rhin, 67401 Illkirch Cedex, France.
Abstract:
Development of novel anticancer formulations is a priority challenge in biomedicine. However, in vitro models based on monolayer cultures (2D) which are currently used for cytotoxicity tests leave much to be desired. More and more attention is focusing on 3D in vitro systems which can better mimic solid tumors. The aim of the study was to develop a novel one-step highly reproducible technique for multicellular tumor spheroid (MTS) formation using synthetic cyclic RGD-peptides, and to demonstrate availability of the spheroids as 3D in vitro model for antitumor drug testing. Cell self-assembly effect induced by addition of both linear and cyclic RGD-peptides directly to monolayer cultures was studied for 12 cell lines of various origins, including tumor cells (e.i. U-87 MG, MCF-7, M-3, HCT-116) and normal cells, in particular L-929, BNL.CL2, HepG2. Cyclo-RGDfK and its modification with triphenylphosphonium cation (TPP), namely cyclo-RGDfK(TPP) in a range of 10-100μM were found to induce spheroid formation. The obtained spheroids were unimodal with mean sizes in a range of 60-120μm depending on cell line and serum content in culture medium. The spheroids were used as 3D in vitro model, in order to evaluate cytotoxicity effects of antitumor drugs (doxorubicin, curcumin, temozolomide). The developed technique could be proposed as a promising tool for in vitro test of novel antitumor drugs.
Insights
Researchers developed a new method using cyclic RGD-peptides to create multicellular tumor spheroids (MTS). These 3D models offer a better in vitro system for testing new anticancer drugs and formulations.
Area of Science:
- Biomedicine
- Oncology
- Drug Discovery
Background:
- Current 2D monolayer cell cultures for cytotoxicity testing have limitations in mimicking solid tumors.
- There is a growing need for advanced 3D in vitro models that better represent tumor microenvironments.
- 3D in vitro systems are crucial for accurate preclinical evaluation of anticancer agents.
Purpose of the Study:
- To develop a novel, reproducible one-step technique for forming multicellular tumor spheroids (MTS).
- To utilize these MTS as a 3D in vitro model for evaluating antitumor drug efficacy.
- To investigate the use of synthetic cyclic RGD-peptides in inducing cell self-assembly for spheroid formation.
Main Methods:
- Studied the cell self-assembly effect of linear and cyclic RGD-peptides on 12 different cell lines (tumor and normal).
- Utilized cyclo-RGDfK and its TPP-modified version (cyclo-RGDfK(TPP)) at concentrations of 10-100μM.
- Characterized the resulting spheroids for unimodality and size (60-120μm) based on cell line and culture conditions.
Main Results:
- Cyclo-RGDfK and cyclo-RGDfK(TPP) effectively induced MTS formation across various cell lines.
- The developed MTS were unimodal and exhibited controlled size ranges.
- The MTS model was successfully used to evaluate the cytotoxicity of doxorubicin, curcumin, and temozolomide.
Conclusions:
- A novel, efficient one-step method for generating MTS using cyclic RGD-peptides was successfully developed.
- The resulting MTS serve as a viable and promising 3D in vitro model for antitumor drug screening.
- This technique offers a valuable tool for advancing the in vitro testing of novel anticancer drugs and formulations.
