Previously cross-linked hydrogels for three dimensional scaffold were made from a mixture of poly(vinyl alcohol)(PVA) and gelatin using a 60Co r-ray irradiation technique. These hydrogels exhibited desirable physical properties as the 3D constructs for diploid cell cultures. The cell adhesion in the hydrogels increased upon increasing the gelatin content. In addition, the cells cultured in gelatin containing hydrogels showed the improved spreading ability and growth ratio. As a continuation study, biocompatibility profiles of gelatin-poly(vinyl alcohol) hydrogels were investigated in vitro and in vivo, in accordance with the ISO 10993-5. Test results were cpmapred to polyurethane(PU) as a positive control and low-density poly ethylene(LDPE) as a negative control.Although the results have small differences with respect to the gelatin contents, all samples showed negligible toxicity to the cells and tissues.