Development and characterization of gelatin-based scaffolds for 3D cancer cell culture

Two-dimensional (2D) cell culture is a commonly utilized method in laboratories for growing and maintaining cells, particularly in cancer research under controlled conditions. A critical factor in cancer cell culture is preserving cell viability and functionality, as cancer cells are highly sensitive to changes in their environment. However, the three-dimensional (3D) structure of tumors cannot be effectively replicated in 2D cell culture. In contrast, 3D cell culture provides significant advantages over traditional 2D systems, primarily by cultivating cancer cells in an environment that more accurately resembles the 3D architecture and complexity of tumors in vivo. This study is designed to develop cancer-specific gelatin-based scaffolds for developing a 3D (spheroid) culture model. These scaffolds were prepared reproducibly using pig skin gelatin, cold-water fish gelatin, and bovine skin gelatin. Phytagel and agarose-based gels (control groups) were used for comparison. The scaffold structures were characterized by Fourier transform infrared (FT-IR) spectroscopy and scanning electron microscopy (SEM). MCF-7, HeLa, and HT-29 cancer cells were seeded on gelatin substrates and imaged by inverted microscopy. FT-IR analysis showed that the scaffolds were successfully prepared, while SEM analysis showed that the scaffolds were highly porous. The cancer cell lines were successfully grown on scaffolds and were shown to aggregate to form spherical structures. In this study, for the first time, 3D structures were generated from monolayer cell structures by using different gelatin structures. Many 3D cell studies will benefit from the fact that the resulting gelatin scaffolds are biocompatible and support infiltration and proliferation.