In Vitro Assessment of the Gene Expression of EZH-2 and P300 During Motor Neuron Differentiation of Human Umbilical Cord Blood Mesenchymal Stem Cells

Introduction: maintenance of neurogenesis depends on the function of some histone-modifying enzymes; including enhancer of zeste homolog 2 (ezh2) and histone acetyltransferases (p300). the mechanism of epigenetic regulation and gene expression underlying the transition of human umbilical cord blood mesenchymal stem cells (hucb-mscs) into mns has not been fully clarified. methods: two morphogens; sonic hedgehog (shh: 100 ng/ml) and retinoic acid (ra: 0.01 mm) were involved in the specification of hucb-mscs into mns after msc characterization using flow cytometry. real time-quantitative pcr and immunocytochemistry were performed to find the expression of the genes at the level of mrna and protein. results: the expression of mn-related markers was confirmed at the level of mrna and protein by induction of differentiation. the results were confirmed by immunocytochemistry and showed those mean cell percentages of 55.33%±15.885% and 49.67%±13.796% could express islet-1 and chat, respectively. the gene expression level of islet-1 and chat was significantly increased in the first and second week of exposure, respectively. after two weeks, the expression level of p300 and ezh-2 genes increased remarkably. no significant expression of mnx-1 was detected when compared to the control sample. conclusion: mn-related markers, islet-1 and chat, were detected in differentiated cells of hucb-mscs, supporting the potency of cord blood cells in the regeneration of mn-related disorders. assessing these epigenetic regulatory genes at the protein level can be suggested to confirm their functional epigenetic modifying effects during motor neuron differentiation.