Differentiation of human olfactory mucosa-derived neural stem cells into dopaminergic neuron-like cells

Title: Differentiation of neural stem cells derived from human olfactory mucosa into dopaminergic neuron-like cells Aim: The aim of this thesis was to isolate neural stem cells from human olfactory mucosa (hOM) by performing in vitro stem cell isolation methods and to differentiate the cultured stem cells into dopaminergic neuron-like cells under the influence of certain neurotrophic factors in vitro by performing cultivation, proliferation and characterization analyses. Methods: Neural stem cells (hO-NCs) were isolated from human olfactory mucosa and immunophenotypic and MTT analyses were performed for their characterization. hO-NCs were cultured in 3D to form neurospheres. Immunophenotypic and neuronal differentiation assays were performed for characterization analyses of neurospheres. Then hO-NSCs were differentiated into dopaminergic neuron-like cells in vitro. After differentiation analysis, immunophenotypic and genotypic characterization analyses of dopaminergic neuron-like cells were performed. Results: Results: Isolation of NSCs from hOM was performed and positive expression of NSCs-specific cell markers (CD56, CD133, SOX2 and NESTIN) was demonstrated. For characterization analyses, neurospheres differentiated in neuronal and glial direction were positively expressed with immunofluorescent labeling of neuronal and glial cell-specific cell markers (TUBB3, GFAP, NG2). Immunocytochemical analysis and RT-PCR analysis were performed for the characterization of hO-NSCs differentiated into dopaminergic neurons in vitro with dopaminergic neuron-specific protein (TH) and gene (Dat, TH, Pitx3, Nurr1). As a result, the positivity of dopaminergic neuron-specific protein (TH) and expression of genes (Dat, TH, Pitx3, Nurr1) were significantly increased. Conclusions: According to the results obtained, it was concluded that a resistant to cell culture, neuronal and glial differentiation, easy to obtain and low-cost neural stem cell source that can be used in cell replacement therapies in neurodegenerative diseases can be created.