Objectives: The purpose of the present in vitro study was to evaluate the biocompatibility of mineral trioxide aggregate (MTA) mixed with glass ionomer cement (GIC), and to compare it with that of MTA, GIC, IRM and SuperEBA. Materials and Methods: Experimental groups were divided into 3 groups such as 1 : 1, 2 : 1, and 1 : 2 groups depending on the mixing ratios of MTA powder and GIC powder. Instead of distilled water, GIC liquid was mixed with the powder. This study was carried out using MG-63 cells derived from human osteosarcoma. They were incubated for 1 day on the surfaces of disc samples and examined by scanning electron microscopy. To evaluate the cytotoxicity of test materials quantitatively, XTT assay was used. The cells were exposed to the extracts and incubated. Cell viability was recorded by measuring the optical density of each test well in reference to controls. Results: The SEM revealed that elongated, dense, and almost confluent cells were observed in the cultures of MTA mixed with GIC, MTA and GIC. On the contrary, cells on the surface of IRM or SuperEBA were round in shape. In XTT assay, cell viability of MTA mixed with GIC group was similar to that of MTA or GIC at all time points. IRM and SuperEBA showed significantly lower cell viability than other groups at all time points (p < 0.05). Conclusions: In this research MTA mixed with GIC showed similar cellular responses as MTA and GIC. It suggests that MTA mixed with GIC has good biocompatibility like MTA and GIC.