title: Enhanced dopamine release by mesenchymal stem cells reprogrammed neuronally by the modulators of SMAD signaling, chromatin modifying enzymes, and cyclic adenosine monophosphate levels.
abstract: Recently, using the chemical genetics approach for cell reprogramming, via the combination of small molecule modulators of chromatin modifying enzymes, specific SMAD signaling pathways, and cyclic adenosine monophosphate levels, we have been able to generate neuronallike cells predominantly positive to mature neuronal and dopaminergic markers. This study aimed to characterize further the dopaminergic properties of neurally induced (NI) human bone marrow-derived mesenchymal stem cells (hMSCs) and to determine whether addition of sonic hedgehog (SHH)/fibroblast growth factor 8 (FGF8) to NI medium could promote further dopaminergic maturation. Dopaminergic differentiation was evaluated by immunocytochemistry, reverse transcription-polymerase chain reaction, Western blot, and enzyme-linked immunosorbent assay. Results demonstrated that release of dopamine by NI-hMSCs differentiated with SMAD inhibitor supplementation significantly increased from picogram to nanogram levels, with a tendency of further increase when supplemented by SHH/FGF8. Direct generation of dopaminergic cells from adult hMSCs by using this reprogramming approach may have significant implications for understanding the mechanism underlying cell plasticity and may open new potentialities for cell replacement therapies.
authors.affiliation: Neuroscience Research Laboratories, Department of Neurosurgery, VA Medical Center, Milwaukee, Wisc.
authors.forename: Ryan T
authors.initials: RT
authors.lastname: Funk
authors.forename: Arshak R
authors.initials: AR
authors.lastname: Alexanian
mesh_terms: D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation; D002478:Cells, Cultured; D003470:Culture Media; D000242:Cyclic AMP / Q000378:metabolism*; D004298:Dopamine / Q000378:metabolism*; D051524:Fibroblast Growth Factor 8 / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D053823:Hedgehog Proteins / Q000494:pharmacology; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D015398:Signal Transduction; D051785:Smad Proteins / Q000378:metabolism
chemical_list: D003470:Culture Media; D053823:Hedgehog Proteins; D051785:Smad Proteins; D051524:Fibroblast Growth Factor 8; D000242:Cyclic AMP; D004298:Dopamine
keywords: -p; -p1; -p2; B3 T; BSA; DA; ELISA; FGF8; NI; RT-PCR; SHH; SYN; bovine serum albumin; cDNA; complementary deoxyribonuclease; dopamine; enzyme-linked immunosorbent assay; fibroblast growth factor 8; hMSCs; human bone marrow-derived mesenchymal stem cells; mouse monoclonal anti-β-III-tubulin; neurally induced; protocol; protocol 1; protocol 2; rabbit polyclonal antisynaptophysin; reverse transcription-polymerase chain reaction; sonic hedgehog