Safety assessment and in vitro cytotoxicity profiling of fruiting bodies from Lentinula edodes (shiitake) strains in HeLa and MCF-7 cell lines

Medicinal mushrooms offer diverse therapeutic benefits, but proper administration is crucial to avoid long-term health issues linked to fungal toxicity. This study evaluated the safety and in vitro cytotoxic potential of Lentinula edodes (shiitake) mushrooms for broader use in food and health supplements. Two strains, DMRO-34 and DMRO-356, were tested for cytotoxic and nutraceutical properties. In vitro analysis using MTT and LDH assays on human cervical (HeLa) and breast cancer (MCF-7) cell lines showed that DMRO-356 exhibited greater anti-proliferative effects compared to DMRO-34. β-glucan content was quantified in both strains using a Megazyme assay as part of the compositional profile. In vivo studies with a limit dose of 5000 mg/kg body weight assessed acute and sub-acute toxicity over 14 and 28 days. Both strains showed no observable adverse effects (NOAEL) in rats, confirming their safety at the tested dose. The results indicate that L. edodes may possess bioactive properties and are well-tolerated in animal models. While the in vitro cytotoxic effects are promising, the in vivo results presented here establish their safety profile; however, further in vivo efficacy investigations are required to validate their therapeutic potential. Overall, these findings suggest that DMRO-34 and DMRO-356 are safe for use and may act as natural therapeutic agents, with DMRO-356 showing greater efficacy. This study supports the safe incorporation of L. edodes mushrooms into dietary supplements and functional foods, and highlights their potential role in adjunct strategies for cancer prevention and immune support.