556 Functional and proliferative melanocyte-derived pluripotent stem cells: from developmental studies to clinical perspectives

Over the last decade, various protocols have been published to generate melanocytes from human pluripotent stem cells (CSP). However, there is limitations in their use specifically for developmental research studies and clinical applications in which a high purity of mel-CSP capable of high proliferative capacity is critical for a large-scale production of cells. To overcome the shortcomings associated to the previous protocols, we have successfully developed an efficient and robust stepwise method to generate a pure and homogenous melanocyte population with higher proliferate rate in culture. A transcriptome analysis in a time-dependent manner was performed providing a spectrum of molecular signatures corresponding to different steps of the melanocyte lineage development. Derived melanocytes exhibit all the melanocytic markers, produce melanin, synthetize melanosomes, and home to the basal layer of the epidermis when incorporated into immunodeficient mice. Further, mel-CSP are capable to proliferate in long-term in culture without evidence of senescence or crisis. Taken together, our results represent a step toward for studying both, normal human development, and molecular mechanisms responsible for genetic diseases. Thus, this unlimited resource of melanocytes provides a promising system to produce in a large-scale up of cells for regenerative medicine.