Differentiation of Mesenchymal Stem Cells Derived From Human Adipose Tissue Into Cholinergic-like Cells; in vitro

Abstract

Cholinergic associated diseases currently are major cause of neurological and neurodegenerative disabilities. As the drugs are not efficient in improving the suffered tissues, stem cell treatment is considered as an effective strategy for substituting the lost cells. In the current study, we set out to investigate the differentiation properties of human adipose-derived mensenchymal stem cells (AD-MSCs) into cholinergic-like cells by two morphogens; including retinoid acid (RA) and Sonic hedgehog (Shh) using a three- step in vitro procedure. The results were evaluated using Real-time PCR, Flowcytometry and Immunocytochemistry for two weeks. Our data showed that the cells could express cholinergic specific markers; including Islet-1, AChE, SMI-32 and Nestin at the level of mRNA and protein. We could also quantitatively evaluate the expression of Islet-1, AChE and Nestin at 14 days post- induction using flowcytometry. It is concluded that human AD-MSCs are potent type cell to differentiate into cholinergic like cells in the presence of RA and Shh through a three- step protocol; thus it could be a suitable cell candidate for regeneration of cholinergic associated diseases; however, more functional and electrophysiological analysis are needed.