Melanocytes are pigment-producing cells generated from neural crest cells (NCCs) that delaminate from the dorsal neural tube. In a wide variety of vertebrate embryos, melanocytes are exclusively derived from NCCs. We show that a NCC population that is not derived from Sox1+ dorsal neuroepithelial cells but are derived from Sox1- cells differentiate into a significant population of melanocytes in the skin of mice. Later, these Sox1- cells clearly segregate from cells that originated from Sox1+dorsal neuroepithelial cell-derived NCCs. The possible derivation of Sox1- cells from epidermal cells also strengthens their non-neuroepithelial origin.