In Vitro Derivation of Melanocytes from Embryonic Neural Crest Stem Cells

Abstract

During development, melanocyte progenitors originate from the neural crest, a transient embryonic structure in vertebrates that gives rise to a variety of cell types including neurons and glia of the peripheral nervous system, smooth muscle cells of the cardiovascular system, chondrocytes and osteoblasts of the craniofacial elements, and pigment cells in the skin. In this chapter, we describe a method for the differentiation of multipotent embryonic neural crest stem cells into differentiated pigmented melanocytes by using in vitro explant culture system. This protocol allows the dissection of genetic and cellular mechanisms regulating neural crest stem cell and melanocyte development. Based on this knowledge it is possible to make predictions about processes that might also be implicated in melanoma initiation and progression. AbstractIn recent years, it has become apparent that the mechanisms by which cancer cells self--renew, migrate and invade, resemble their embryonic counterparts and that during the steps of the initiation and progression cancer cells can reprogram into embryonic--like cells (1, 2). Melanoma is one of the most aggressive human cancers and it is believed to originate from the genetic deregulation of the cells of the melanocytic lineage. As many other cancers, melanoma cells display the expression of developmental genes and can be reprogrammed when exposed to embryonic environment (3, 4). During development, melanocyte progenitors originate from the neural crest, a transient embryonic structure in vertebrates that gives rise to a variety of cell types including neurons and glia of the peripheral nervous system, smooth muscle cells of the cardiovascular system, chondrocytes and osteoblasts of the craniofacial elements, and pigment cells in the skin. In this chapter, we describe a method for the differentiation of embryonic neural crest stem cells into melanocytes. This protocol allows the dissection of genetic and cellular mechanisms regulating neural crest stem cell and melanocyte development. Based on this knowledge it is possible to make predictions about processes that might also be implicated in melanoma initiation and progression.

Wnt Signaling in Neural Crest Ontogenesis and Oncogenesis

“…Several studies have been performed in vivo to address the developmental potential of individual NC cells by retrovirus-mediated gene transfer or dye injection to label cell lineages [196,197,198]. These studies showed that at least some NC cells generate different cell types in vivo, including smooth muscle cells, neurons, glia, and melanocytes [198,199]. Cultured NC cells from mouse neural tube explants derived prior to NC cell migration demonstrated their multipotency in vitro [200].…”

Stem Cell-Derived Models of Neural Crest Are Essential to Understand Melanoma Progression and Therapy Resistance

“…In the mouse system, melanocyte generation from embryonic NCSC has been performed by addition of SCF and END3 in the culture medium (Shakhova and Sommer, 2015). In the human system, protocols of melanocyte differentiation from PSCs through a NC stage have also been described.…”

Prospect of Human Pluripotent Stem Cell‐Derived Neural Crest Stem Cells in Clinical Application

“…Shakhova and Sommer [84] have established a differentiation approach to generate melanocytes from embryonic NCSCs in 10 days by the addition of a cocktail of specific growth factors such as ET-3 and mouse SCF. Their study showed that after 10-day differentiation, the majority of NCSCs presented a melanocytic phenotype indicating the in vitro differentiation ability of NCSCs into melanocytes.…”