Lentinus tigrinus is a significant edible and medicinal mushroom. It was collected from University of the Punjab, Lahore under a Morus tree. For the domestication of this wild significant species, its three parameters i.e., culturability, spawn production and cultivation potential were assessed using different synthetic culture media and substrates. Completely randomized design was used to determine the parameters and all the effects were evaluated in triplicates. Among these different media used, maximum mycelial growth where observed on the potato dextrose agar (PDA) at 35°C, followed by the compost extract agar (CEA) medium, malt extract agar (MEA), glucose peptone agar (GPA), and Saboraud dextrose agar (SDA). Spawning material was prepared on rice, sorghum and barley grains by the cultured mycelium on PDA medium. Sorghum grains were found as the appropriate medium for spawn production of this fungus. Cultivation potential and biological efficiency was assessed on the different substrates i.e., pure rice straw, pure sawdust, pure tea-waste, mixture of sawdust and rice straw, mixture of tea waste and sawdust and mixture of tea waste and rice straw. Mixed substrate of sawdust and rice straw at 30oC showed the maximum yield. Tea-waste medium was used as the casing material and proved very effective. These results indicated that Lentinus tigrinus exhibit the growth potential and its domestication can compete with nutritional and medicinal peculiarities of one of the most cultivated species, L. edodes.