Establishment of Culture System Capable of Inducing Neural Crest Cells from ES Cells

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Mouse embryonic stem (ES) cells are totipotent, primitive, ectodermal-like cells derived from inner cell mass of 3,5 day blastocysts. As an in vitro model for melanocyte differentiation, we have developed the culture system to induce melanocytes from totipotent ES cell line. When undifferentiated ES cells were co-cultured with a bone marrow-derived stromal cell line, a very small but significant number of melanocytes were reproducibly generated. This process was greatly enhanced by addition of dexamethasone to the culture and strictly depended on the c-Kit signaling and endothelin 3 signaling. In this study, we found that caudalizing factor retinoic acid enhanced the efficiency of melanocyte induction in this culture. The early precursors for melanocytes, which proliferate clonally after several passages, were efficiently induced in the culture and their self-renew was stimulated by the addition of endothelin 3 and also by retinoic acid. These results suggests that neural crest cells are induced in this culture system upon corresponding in vivo development.