P64. Isolation and characterization of neural crest stem cells from adult human hair follicles

Abstract

Neural crest (NC) is a transient embryonic tissue, whose cells are motile and multipotent until they reach their destination and differentiate according to microenvironmental cues into variety of cell types. However, a subpopulation of these cells remains multipotent. They were found, among other locations, in a bulge of adult murine whisker follicle and were designated epidermal neural crest stem cells (EPI-NCSCs). The aim of this work is to ascertain whether the EPI-NCSCs could be isolated from human hair follicles as well. Due to their exceptional properties, they could represent potential candidate for stem cell therapy. The presented work focuses on the isolation and characterization of EPI-NCSCs from human skin. We obtained population of cells that expressed markers of NC, NC progeny and general stem cell markers. After prolonged cultivation, subpopulation of cells spontaneously differentiated into some of NC derivatives, i.e. neurons, smooth muscle cells and Schwann cell progenitors. Targeted differentiation with neuregulin-1 highly increased the number of Schwann cells in culture. Human EPI-NCSCs were also able to grow under non-adherent conditions and form three-dimensional spheres. Microarray analysis was performed and gene profile of human EPI-NCSCs was compared with the list of key genes of murine EPI-NCSCs, and the list of genes upregulated in newly induced NC cells. This revealed 94% and 88% similarity, respectively. All the presented results strongly support the NCSCs identity and multipotency of isolated human cells. These cells thus could be a good candidate for stem cell therapy, especially because of the easy accessibility of donor tissue.