Induction of mesenchymal stem cell-like transformation in rat primary glial cells using hypoxia, mild hypothermia and growth factors

The transformation of rat primary glial cells into mesenchymal stem cells (MSCs) is intriguing as more seed cells can be harvested. The present study aimed to evaluate the effects of growth factors, hypoxia and mild hypothermia on the transformation of primary glial cells into MSCs. Rat primary glial cells were induced to differentiate by treatment with hypoxia, mild hypothermia and basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF). Immunohistochemistry and western blotting were then used to determine the expression levels of glial fibrillary acidic protein (GFAP), nestin, musashi-1, neuron specific enolase (NSE) and neuronal nuclei (NeuN), in each treatment group. bFGF and EGF increased the proportion of CD44(+) and CD105(+) cells, while anaerobic mild hypothermia increased the proportion of CD90(+) cells. The combination of bFGF and EGF, and anaerobic mild hypothermia increased the proportion of CD29(+) cells and significantly decreased the proportions of GFAP(+) cells and NSE(+) cells. Treatment of primary glial cells with bFGF and EGF increased the expression levels of nestin, Musashi-1, NSE and NeuN. Anaerobic mild hypothermia increased the expression levels of Musashi-1 and decreased the expression levels of NSE and NeuN in glial cells. The results of the present study demonstrated that bFGF, EGF and anaerobic mild hypothermia treatments may promote the transformation of glial cells into MSC-like cells, and that the combination of these two treatments may have the optimal effect.