(176b) Neural Crest Stem Cells from Human Epidermis Skin Tissue

Neural crest Stem Cells (NCs) are transient cells generated during early vertebrate development. These multipotent cells show extensive migratory and proliferative capacity as well as the potential to differentiate into multiple lineages from craniofacial skeletal tissues to cells of the peripheral nervous system. However, clinical application of these cells is hindered by the limited availability of cell sources. Previously we showed that NC can be derived from neonatal keratinocytes (KC) with a differentiation protocol that did not involve genetic modification. Although a very promising result, it was not clear whether NC could also be derived from the skin of adult donors, who are most likely in need of cellular therapies. Here we showed that NCs can be obtained from epidermal KC of human skin tissues of adult donors ranging from 65 to 97 years of age. Adult NCs derived from KC expressed key NC markers including lineage-specific transcription factors such as SOX10, FOXD3 and intermediate filament, NES. Notably, adult NC did not exhibit the hallmarks of cellular senescence (aging) as evidenced by examination of multiple markers of cellular senescence including p16, p21, p53, oxidative DNA damage or SA-b-galactosidase expression. Using protein expression and functional assays, we demonstrated that KC-derived NC could be coaxed to differentiate into all NC-specific lineages including neurons, Schwann cells, melanocytes, and smooth muscle cells using appropriate differentiation strategies. Finally, implantation of adult NC in chick embryos showed that these cells contributed to the full repertoire of NC derivatives, from neural and glial cells to mesenchymal and pigment cells. Our results suggest that multipotent NC stem cells can be derived from adult skin, without genetic modification or reprogramming to the pluripotent state. These findings may have significant implications for treatment of neurodegenerative diseases for which cell sourcing remains a major roadblock to development of cellular therapies.