Embryonic stem (ES) cell lines have provided very useful models to analyse differentiation processes. We present here the development of a differentiation system using ES-like cell lines from medaka. These cells were transfected with the melanocyte specific isoform of the microphtalmia-related transcription factor (Mitf). Mitf is a basic helix-loop-helix-leucine zipper transcription factor whose M isoform is restricted to neural crest derived melanocytes and is essential for the development of these cells in vertebrates from mammals to fish. What is not clear yet is whether Mitf is a downstream factor or a master regulator of melanocyte commitment and differentiation. Expression of Mitf in the ES-like cells from medaka led to the induction of cells that, by morphology, physiology and gene expression pattern, were confirmed to be fully differentiated pigment cells. Mitf expression is therefore sufficient for the proper differentiation of medaka pluripotent stem cells into melanocytes.