When conventional eye drops are applied topically, the bioavailability of the drug in the eye is very low due to the rapid elimination of the active substance. The main characteristic of in situ gelling systems is the prolongation of the retention time on the surface of the eye and consequently the increase in the bioavailability of the drug. The aim of this diploma thesis was to investigate the suitability of in vitro corneal epithelial models for the evaluation of the biocompatibility of in situ gelling system that are based on poloxamers and chitosan. For this purpose, the immortalized corneal epithelial cell line, HCE-T, and an in situ gel system consisting of P407 (14,2%, m/m), P188 (17%, m/m) and chitosan (0,25%, m/m) were used. The cytotoxicity of the in situ gelling system was investigated on a monolayer of a human corneal epithelial cell line, HCE-T and on in vitro corneal models based on bilayer and multilayer of HCE-T cells. After treating cells with the formulation, cell viability was evaluated by MTT test. The cytotoxic effect was not observed after a monolayer of HCE-T cells was exposed to the in situ gel system for 5, 10 and 15 minutes. The in vitro corneal epithealial model enabled the determination of the influence of chitosan present in the in situ gel system on the opening of tight junctions between the corneal epithelial cells by measuring the transepithelial electric resistance. The cytotoxic effect was not observed during the 30-minute exposure of the in vitro corneal epithelial models to the in situ gelling system. In vitro corneal epithelial models proved less sensitive in comparison to monolayer cells and are, therefore, more suitable models for studying cytotoxicity of the in situ gelling formulations.