Abstract

Alginate bead-based three-dimensional (3D) culture provides a biomimetic environment that closely mimics in vivo conditions, thereby effectively maintaining the morphology and cellular connections of HepG2 cells. During the bead formation process, an electrospray system was used to vary the voltage provided to the nozzle to control the size and homogeneity of the beads. Alginate beads formed at 3 kV exhibited the highest cell viability, exceeding 80%, with an average bead diameter of approximately 2–3 mm. Cytotoxicity testing using Thymoquinone (TQ) revealed that HepG2 cells cultured in a 3D alginate system exhibited significantly greater resistance compared to those cultured in a two-dimensional (2D) monolayer. The concentration of TQ required to inhibit 50% of cell viability (IC50) was 916 μg/ml in 3D culture, whereas only 384 μg/ml was required in 2D culture. The 3D alginate bead model enhances the accuracy of drug sensitivity evaluation and more closely mimics in vivo tumor behavior. This approach offers a viable alternative to conventional cell culture models and may reduce the need for animal testing in preclinical research.

Key words: Alginate beads; HepG2 cell lines; Three-dimensional culture; Thymoquinone; Electrospray technique; Cytotoxicity assay.