Vitiligo also known as Leucoderma is caused by the loss of pigment, resulting in irregular pale patches of skin. Vitiligo develops patches of depigmented skin appearing on extremities. The objective of the present study was to evaluate the cytotoxicity and in-vitro melanogenic activity on bark of Dalbergia sissoo Roxb (Fabaceae). The various successive bark  extracts have been individually evaluated for trials of spontaneous melanin content, and cell viability by the MTT assay in murine B16F10 melanoma cells in-vitro. Based on the percentage of cell viability assay, graded concentration of extracts were taken for invitro melanogenic activity. The result indicated that Ethyl acetate extract of bark of Dalbergia sissoo was found to be non-toxic and increased melanin activity as compared to Hexane and Ethanol extracts. From the above result, it can be concluded from this study that the bark of Dalbergia sissoo stimulates B16F10 melanogenesis at very low concentrations. These findings support the folk medicinal use of Dalbergia sissoo on the treatment of hypopigmentation diseases, such as vitiligo.