Differentiation of human mesenchymal stem cells through the natural matrix to neurospheres for cholinergic-like cells

Abstract

Abstract Background In Alzheimer's Disease there is an impairment of the cholinergic system, causing loss of neurons, impairment of intellectual abilities. In this context, the mesenchymal stem cells (MSC) and its applications in cell therapies become target of the research, which may contribute to the treatment of neurodegenerative diseases. Through the differentiation potential of MSCs, prospecting functional repair of the injured tissue from cholinergic neuronal cells could be a potential treatment. The aims was to evaluate the possibility of differentiation of MSCs from the human umbilical cord in nestin-positive neural precursor cells (NPCN+) through the NFBX into cholinergic ‘like’ cells. Methods The isolation of hMSCs from Wharton's jelly (WJ) was by the explant and mononuclear cells by density gradient. hMSCs were plating in natural matrix as NFBX for neurospheres production. Neural precursor cells were subjected to standard cholinergic differentiation protocol. Dissociated neurospheres, neural precursor cells and cholinergic-like cells were characterized by immunocytochemistry. The RT-PCR was done. Results hMSCs were CD73+, CD90+, CD105+, CD34- and CD45- and demonstrated the trilineage differentiation. Neurospheres and their isolated cells were nestin positive, and also expressed NESTIN, MAP2, ßIII-TUBULIN, GFAPgenes. Neural precursor cells that were differentiated in cholinergic-like cells expressed ßIII-TUBULIN protein and choline acetyltransferase enzyme. Conclusions hMSCs on the natural matrix were capable of differentiating hMSC into neurospheres, obtaining neural precursor cells without growth factors or gene transfection before cholinergic differentiation.