Isolation, culture and neurogenic differentiation of human dental stem cells

A Dissertation submitted to the Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, in fulfillment of the requirements for the degree Of Master of Science in Medicine, 2016.Dental stem cells (DSCs) have been identified in teeth and their supporting tissues. They represent an exclusive source of adult stem cells, easily isolated and manipulated for tissue repair and regeneration. This research project evaluated the neurogenic potential of the dental pulp stem cells (DPSCs) and stem cells from the pulp of human exfoliated deciduous teeth (SHEDs) in a South African cohort. Sixty non-carious permanent and deciduous teeth were extracted from healthy patients aged between 18 and 30 years and 5 and 10 years, at the University of the Witwatersrand's Oral Health Clinic in Johannesburg Charlotte Maxeke Academic Hospital, South Africa. The cells, isolated from the extracted pulp tissue were cultured, counted and then phenotyped by flow cytometry analysis. The cells were further expanded in a neural induction medium and immunocytochemistry analysis for Ki-67, doublecortin (DCX) and nestin were performed. Large colonies of both DPSCs and SHEDS were harvested from the extracted pulp tissues and positively cultured. Flow cytometry analysis confirmed the presence of CD44+ and CD29+ cells as well as the known mesenchymal stem cell markers CD90 and CD105. Both DPSCs and SHEDs demonstrated successful proliferation and neural differentiation. This study confirmed that DPSCs and SHEDs are highly proliferative human adult stem cells that exhibit a neurogenic potential that may contribute in the treatment of neurological disorders.AC201