Agaricus blazei, Grifola frondosa and Hericium erinaceus are popular medicinalmushroom. They have beenwidely consumed inmany countries, including Malaysia, due to its therapeutic properties and commonly used as traditional medicine. This study was carried out to determine the in vitro toxicity of these medicinal mushrooms that were cultivated in Malaysia and their possible risk to human health. The cytotoxic and genotoxic activity were evaluated against Chinese hamster fibroblast cell line (V79-4) using MTT assay and alkaline comet assay. Five different concentrations of the extracts (2mg/ml, 1mg/ml, 0.5mg/ml, 0.25mg/ml and 0.125mg/ml) in 70% methanol and aqueous solvent were used. The cytotoxicity was determined following 24 hours treatment. Our results demonstrated that all extracts show decreased in cell viability at the maximumconcentration of 2mg/ml. On the other hand, none of the extracts caused DNA damage to the V79-4 cells at 2mg/ml following 24 hour incubation, as compared to the positive control, menadione (tail moment- 13.135±2.273). In conclusion, several mushroom extracts showed weak cytotoxic effect but did not cause DNA damage against Chinese hamster fibroblast cell line (V79-4), suggesting that these mushrooms are safe to consume.