Two-Step Induction of Dopaminergic Neurone Differentiation of Leukapharesis-Derived Mesenchymal Stem Cells

Abstract

Abstract:Background: Neurodegenerative diseases represent a great medical challenge with marked consequences on quality of life of the patients and their families. Parkinson's disease (PD) constitutes a relatively common neurodegenerative disorder characterized by defect in dopaminergic neurons. The regenerative effect of mesenchymal stem cells (MSCs) stimulated research into their effect in treatment of PD. Subjects and Methods: Leukapharesis-derived MSC were isolated from 10 leukapharesis products using plastic adherence. Isolated MSCs were passaged, and passage III cells were induced to dopaminergic neurones using two-step protocol applying sequentially nerve growth factor and addition of ascorbic acid. Neuronal induction was evaluated by immunostaining against neurofilament protein, while dopaminergic neurons induction was evaluated using immunostaining against tyrosine hydroxylase. Results and Conclusions: MSCs were isolated in a rate of 0.12%-0.15% of leukapharesis cells, with viability ranging from 70-96%. NF positivity was 15.38±3.84, while the percentage of cells stained for TH was 5.94±0.65.MSCs could be successfully induced to dopaminergic neuron-like cells in vitro.