Abstract

Therapeutic Potential of Human Neural Crest-Derived Stem Cells

Darius Widera, Lecturer, University Of Reading

Restoring the tissue integrity after acute injuries and compensation for the progressive loss of cells during degenerative disorders using current pharmacological treatment strategies is limited and remains challenging. Here, stem cell-based cell replacement therapies represent a promising therapeutic alternative. Although the use of pluripotent stem cells is a possible therapeutic option, the clinical application of foetal tissue, human embryonic stem cells and induced pluripotent stem cells is limited due to solid ethical and practical considerations (e.g. limited availability or tumorigenicity). In contrast, adult neural crest-derived stem cells (NCSCs) can be readily isolated and possess an extraordinarily high developmental potential surpassed only by pluripotent cells. We previously reported isolation of NCSCs from various human craniofacial tissues including the periodontal ligament, hard palate and nasal turbinates. Revealing their multipotent character, such craniofacial NCSCs were able to give rise to mesodermal cells including adipocytes, chondrocytes and osteoblasts in addition to ectodermal derivatives including neurons and glia. Assessing their capability to give rise to mature neurons in the first place, we differentiated human NCSCs towards forebrain-like neurons in vitro, as shown by characteristic morphology and expression of mature neuronal markers including Synaptophysin, vGlut2 and GAT-1. Functionality of NCSC-derived neurons was demonstrated by their ability to recycle synaptic vesicles and repeated calcium spiking after KCl-stimulation. Transplantation into murine organotypic hippocampal slice cultures resulted in neuronal differentiation and integration of NCSCs into pre-existing neuronal networks. In addition, using a protocol based on dual-SMAD inhibition and sequential treatment with SHH and FGF-8 we successfully differentiated NCSCs into TH-positive, dopaminergic-like neurons in vitro. Transplantation of undifferentiated and NCSCs pre-di