Induction of neural differentiation of adipose stromal cells

Adipose stromal cells (ASCs) are progenitor cells capable to differentiate into a large variety of cell types including neuronal cells. Many active ingredients were suggested for the induction of neural differentiation of stromal progenitor cells. However the combination of pharmaceutically approved agents allowing stable induction of neural differentiation of ASCs is not established. Here, we tested the ability of brain derived neurotrophic factor (BDNF) and retinoic acid (RA) alone as well as together with DNA demethylating agent 5-azacytidine to induce stable neural differentiation of ASCs. ASCs were isolated from human or mouse (BI6 strain) sub cutis as described by Zuk et al. At passages 2—5 the cells were induced with NIM (DMEM/F12, 3% FBS and 1 mcM azacytidine supplemented with ImcM RA or 20ng/ml BDNF) for 3 days. The efficiency of neural differentiation was estimated by the change of expression of neuronal markers, including nest in, tubuiin-beta3, neuron-specific enolase 2 and microtubule- associated protein 2, at 3 and 7 days after induction using Real Time PCR. The expression of marker genes increased Б-10 times after incubation in the NIM, supplemented with RA, and up to 4 times in the medium containing BDNF 3 days after induction. Furthermore, ASCs primed to neural differentiation demonstrate significantly better surviving and incorporation in the brain tissue after transplantation into the mouse brain. Taken together, our data suggest that the combination of BDNF or RA with 5-azacytidine could be suggested for the induction of stable neural transdifferentiation.