MSCs can be induced to differentiate into NSC-like cells and exhibit multipotency by generating progenies of different neural lineages in vitro. (A) During differentiation, MSCs develop the properties of neural stem cells (NSCs). The induced cells were observed in the bright fields after 0.5 days and after 14 days. The growth curves of the MSCs and the NSC-like cells, which were obtained by enumerating the cells at each time point under a haemocytometer, are shown (n=3; the calculated doubling times were 20 hours and 40 hours, respectively). (B) After induction, the NSC-like cells were fixed, and immunofluorescence was used to detect the expression of Nestin and Tuj-1. The cells were counterstained using DAPI. The scale bar is 10 μm. To assess the induction efficiency, the cells were co-labelled with antibodies against Nestin and Tuj-1, Sox-2 or Sox1, and the proportions of double-positive cells were 66.83%, 90.92% and 64.71%, respectively. The tests were repeated three times. (C) To determine whether the induced NSC-like cells could differentiate into both neurons and glia cells, MSCs were induced for 14 days under the same conditions and stained using antibodies against neuron- and glial cell-specific proteins. The scale bar is 10 μm.