A GEL CASTING TOOL FOR FABRICATION OF THREE DIMENSIONAL SANDWICH IN-VITRO CULTURE SYSTEM FOR CYTOCOMPATIBILITY EVALUATION OF HYDROGELS

Two-dimensional (2D) culture systems are indispensably used as in-vitro tools to improve our perception and understanding of cell biology, tissue engineering, tissue morphology, mechanisms of diseases, and drug action. However, the conventional 2D culture systems suffer from limitations such as alterations in cell morphology and lack of interactions between extracellular and cellular environments. As a result, three-dimensional (3D) cell cultures have received remarkable attention in studies such as drug discovery and development; the sandwich model being one of the unique cellular models widely used for analyzing tissue metabolism and functions. In this study, we devised a novel Gel Casting Tool (GCT) for in-vitro toxicity analysis. Liver cells need complex 3D micro-architecture to express tissue-specific functions and were suitable for this study. Human Carcinoma Cell line (HepG2 cells) with Gelatin methacrylamide (GelMA) hydrogel was used to check the efficacy of the GCT. The results showed that GCT Sandwiched HepG2 exhibited high cell viability, proliferation, and tissue-specific functions such as albumin and urea over 7 days. The major advantage of this culture model is the maximized availability of cells for observation under microscope and imaging cells in one plane. This culture system has potential application in in-vitro toxicity testing, early drug screening, and evaluation of scaffolds for tissue engineering.