Cholinergic neuronal differentiation of bone marrow mesenchymal stem cells in rhesus monkeys

The purpose of the present study was to determine the best cholinergic neuronal differentiation method of rhesus monkey bone marrow mesenchymal stem cells (BMSCs). Four methods were used to induce differentiation, and the groups were assigned accordingly: basal inducing group (culture media, bFGF, and forskolin); SHH inducing group (SHH, inducing group); RA inducing group (RA, basal inducing group); and SHH+RA inducing group (SHH, RA, and basal inducing group). All groups displayed neuronal morphology and increased expression of nestin and neuron-specific enolase. The basal inducing group did not express synapsin, and cells from the SHH inducing group did not exhibit neuronal resting membrane potential. In contrast, results demonstrated that BMSCs from the RA and SHH+RA inducing groups exhibited neuronal resting membrane potential, and cells from the SHH+RA inducing group expressed higher levels of synapsin and acetylcholine. In conclusion, the induction of cholinergic differentiation through SHH+RA was determined to be superior to the other methods.