Alginate-based 3D cell culture technique to evaluate the half-maximal inhibitory concentration: an in vitro model of anticancer drug study for anaplastic thyroid carcinoma

Hilda Samimi,Parviz Fallah,Alireza Naderi Sohi,Mojdeh Mahdiannasser,Ehsan Arefian,Vahid Haghpanah,Shiva Irani

doi:10.1186/s13044-021-00118-w

Abstract

BackgroundThree-dimensional (3D) cell culture methods are identified for simulating the biological microenvironment and demonstrating more similarity to in vivo circumstances. Anaplastic thyroid carcinoma (ATC) is a lethal endocrine malignancy. Despite different treatment approaches, no improvement in the survival rate of the patients has been shown. In this study, we used the 3D in vitro ATC model to investigate the cytotoxic effect of BI-847325 anticancer drug in two-dimensional (2D)- and 3D- cultured cells.MethodsHuman ATC cell lines, C643 and SW1736, were cultured in one percentage (w/v) sodium alginate. Spheroids were incubated in medium for one week. The reproducibility of the fabrication of alginate beads was evaluated. Encapsulation of the cells in alginate was examined by DAPI (4′,6-diamidino-2-phenylindole) staining. Survival of alginate-encapsulated cells was evaluated by CFSE (5,6-Carboxyfluorescein N-hydroxysuccinimidyl ester) staining. The population doubling times of C643 and SW1736 cell lines cultured in 2D monolayer as well as in 3D system were calculated. The cytotoxic effect of BI-847325 on 2D- and 3D- cultured cell lines was assessed for 24–72 h by MTT [3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide] assay. Finally, the 3D culture results were compared with the 2D culture method.ResultsThe half-maximal inhibitory concentration (IC50) values of BI-847325 were higher in 3D culture compared to 2D culture. The cytotoxicity data indicated that 3D in vitro models were more resistant to chemotherapy agents.ConclusionsThe findings of this study are beneficial for developing in vitro ATC 3D models to analyze the efficacy of different chemotherapy drugs and formulations.

Highlights

Thyroid cancer is the most frequent endocrine malignancy with the highest growing incidence rate between the other types of solid tumors in the United States [1,2,3]
Determination of half-maximal inhibitory concentration (IC50) in the two dimensional (2D) cell culture system Evidence has shown that through carefully selecting thyroid cancer cell lines, an appropriate in vitro model can be developed to analyze the signaling pathways associated with thyroid carcinogenesis [33]
The treatment panel of the cell lines with BI-847325 (0.125–8 μM) for 24-72 h prevented proliferation in two cell lines. In both cell lines, IC50 remained constant after 48 h and IC50 values of BI-847325 on C643 and SW1736 were at 2 μM and 4 μM, respectively (Table 1)

Summary

Introduction

Thyroid cancer is the most frequent endocrine malignancy with the highest growing incidence rate between the other types of solid tumors in the United States [1,2,3]. Cancer investigators rely on 2D cell culture in vitro experiments and animal models to understand the complex biological processes of tumor formation, progression and, its treatment [8, 9]. 3D cell culture models more closely mimic key factors of natural tumor microenvironment such as molecular concentration gradients and crucial cellular processes such as cell to cell and cell to extracellular matrix (ECM) interactions. These culture systems act as a surrogate for animal models and are less expensive and provide quicker results compared for animal models [10,11,12]. We used the 3D in vitro ATC model to investigate the cytotoxic effect of BI-847325 anticancer drug in two-dimensional (2D)- and 3D- cultured cells

Methods

Results

Conclusion