In Vitro Differentiation of Neural Stem Cells into Noradrenergic-Like Cells

Neural stem cells (NSCs) as a heterogeneous multipotent and self- renewal population are found in different
areas in the developing mammalian nervous system, as well as the sub-ventricular zone (SVZ) and the
hippocampus of the adult brain. NSCs can give rise to neurons, astrocytes and oligodendrocytes. The aim of this
study was to differentiate neural stem cells into noradrenergic–like cells in vitro. Neural stem cells were
harvested from SVZ of newborn rat brains. The cells were cultured in DMEM12, B-27 supplemented with 20 ng/
ml (hFGF) and 20 ng/ ml (EGF) for 2 weeks. Neurospheres were differentiated in neurobasal medium, B-27
supplemented with BDNF (50 ng/ ml) and GDNF (30 ng/ ml) for 3 and 5 days. Cell culture techniques and
immunocytochemistry were applied to examine neurospheres and tyrosine hydroxylase positive cells. The
number of neurites was counted 3 and 5 days after the induction of differentiation. Nestin and Sox2 were
expressed in NSCs and neurospheres. NSCs were differentiated into noradrenergic- like cells (NACs). Tyrosine
hydroxylase was detected in these cells. The results of NSCs differentiation for 5 days culture had a significant
decrease (P≤0.05) in the number of TH positive cells with one or two neurite per cell, and a significant increase
(P≤0.05) in the number of TH positive cells with three, four or more neurites per cell, compared with 3 days
culture. Based on these results, NSCs have the ability to differentiate into noradrenergic cells in the presence of
BDNF and GDNF growth factors.