title: Enhancing the efficiency of direct reprogramming of human mesenchymal stem cells into mature neuronal-like cells with the combination of small molecule modulators of chromatin modifying enzymes, SMAD signaling and cyclic adenosine monophosphate levels.
abstract: Advances in cell reprogramming technologies to generate patient-specific cells of a desired type will revolutionize the field of regenerative medicine. While several cell reprogramming methods have been developed over the last decades, the majority of these technologies require the exposure of cell nuclei to reprogramming large molecules via transfection, transduction, cell fusion or nuclear transfer. This raises several technical, safety and ethical issues. Chemical genetics is an alternative approach for cell reprogramming that uses small, cell membrane penetrable substances to regulate multiple cellular processes including cell plasticity. Recently, using the combination of small molecules that are involved in the regulation chromatin structure and function and agents that favor neural differentiation we have been able to generate neural-like cells from human mesenchymal stem cells. In this study, to improve the efficiency of neuronal differentiation and maturation, two specific inhibitors of SMAD signaling (SMAD1/3 and SMAD3/5/8) that play an important role in neuronal differentiation of embryonic stem cells, were added to our previous neural induction recipe. Results demonstrated that human mesenchymal stem cells grown in this culture conditions exhibited higher expression of several mature neuronal genes, formed synapse-like structures and exerted electrophysiological properties of differentiating neural stem cells. Thus, an efficient method for production of mature neuronal-like cells from human adult bone marrow derived mesenchymal stem cells has been developed. We concluded that specific combinations of small molecules that target specific cell signaling pathways and chromatin modifying enzymes could be a promising approach for manipulation of adult stem cell plasticity.
authors.affiliation: Neuroscience Research Laboratories, Department of Neurosurgery, VA Medical Center-Research 151, Milwaukee, WI 53295, United States. aalexan@mcw.edu
authors.forename: Arshak R
authors.initials: AR
authors.lastname: Alexanian
authors.forename: Qing-song
authors.initials: QS
authors.lastname: Liu
authors.forename: Zhiying
authors.initials: Z
authors.lastname: Zhang
mesh_terms: D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D065150:Cellular Reprogramming / Q000187:drug effects*; D042002:Chromatin Assembly and Disassembly / Q000187:drug effects*; D000242:Cyclic AMP / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D015640:Ion Channel Gating / Q000187:drug effects; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects; D051785:Smad Proteins / Q000378:metabolism*; D054852:Small Molecule Libraries / Q000494:pharmacology*; D012964:Sodium / Q000378:metabolism; D013569:Synapses / Q000378:metabolism
chemical_list: D051785:Smad Proteins; D054852:Small Molecule Libraries; D049452:Green Fluorescent Proteins; D012964:Sodium; D000242:Cyclic AMP; D002118:Calcium
keywords: Epigenetics; Mesenchymal; Neuronal; Reprogramming; Stem cells