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...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Iran University of Medical Sciences
2022-09-01
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| Series: | Basic and Clinical Neuroscience |
| Subjects: | |
| Online Access: | http://bcn.iums.ac.ir/article-1-1984-en.html |
| _version_ | 1797285813319368704 |
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| author | Marjaneh Motaghed Davood Sanooghi Zohreh Bagher Faezeh Faghihi Abolfazl Lotfi Ali Shahbazi Mohammad Taghi Jogataei |
| author_facet | Marjaneh Motaghed Davood Sanooghi Zohreh Bagher Faezeh Faghihi Abolfazl Lotfi Ali Shahbazi Mohammad Taghi Jogataei |
| author_sort | Marjaneh Motaghed |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-07T18:08:47Z |
| format | Article |
| id | doaj.art-9e6b45548eb44ddca04204fd1410fe25 |
| institution | Directory Open Access Journal |
| issn | 2008-126X 2228-7442 |
| language | English |
| last_indexed | 2024-03-07T18:08:47Z |
| publishDate | 2022-09-01 |
| publisher | Iran University of Medical Sciences |
| record_format | Article |
| series | Basic and Clinical Neuroscience |
| spelling | doaj.art-9e6b45548eb44ddca04204fd1410fe252024-03-02T07:55:09ZengIran University of Medical SciencesBasic and Clinical Neuroscience2008-126X2228-74422022-09-01135709718In Vitro Assessment of the Gene Expression of EZH-2 and P300 During Motor Neuron Differentiation of Human Umbilical Cord Blood Mesenchymal Stem CellsMarjaneh Motaghed0Davood Sanooghi1Zohreh Bagher2Faezeh Faghihi3Abolfazl Lotfi4Ali Shahbazi5Mohammad Taghi Jogataei6 Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran. Department of Genetics, School of Biological Sciences, Shahid Beheshti University, Tehran, Iran. ENT and Head and Neck Research Center and Department, Five Senses Institute, Hazrat Rasoul Akram Hospital, Iran University of Medical Sciences, Tehran, Iran. Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran. Damavand Agricultural College, Technical and Vocational University, Tehran, Iran. Department of Neuroscience, School of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran. Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran. 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.http://bcn.iums.ac.ir/article-1-1984-en.htmlcord bloodmotor neuronp300ezh-2 |
| spellingShingle | Marjaneh Motaghed Davood Sanooghi Zohreh Bagher Faezeh Faghihi Abolfazl Lotfi Ali Shahbazi Mohammad Taghi Jogataei In Vitro Assessment of the Gene Expression of EZH-2 and P300 During Motor Neuron Differentiation of Human Umbilical Cord Blood Mesenchymal Stem Cells Basic and Clinical Neuroscience cord blood motor neuron p300 ezh-2 |
| title | In Vitro Assessment of the Gene Expression of EZH-2 and P300 During Motor Neuron Differentiation of Human Umbilical Cord Blood Mesenchymal Stem Cells |
| title_full | In Vitro Assessment of the Gene Expression of EZH-2 and P300 During Motor Neuron Differentiation of Human Umbilical Cord Blood Mesenchymal Stem Cells |
| title_fullStr | In Vitro Assessment of the Gene Expression of EZH-2 and P300 During Motor Neuron Differentiation of Human Umbilical Cord Blood Mesenchymal Stem Cells |
| title_full_unstemmed | In Vitro Assessment of the Gene Expression of EZH-2 and P300 During Motor Neuron Differentiation of Human Umbilical Cord Blood Mesenchymal Stem Cells |
| title_short | In Vitro Assessment of the Gene Expression of EZH-2 and P300 During Motor Neuron Differentiation of Human Umbilical Cord Blood Mesenchymal Stem Cells |
| title_sort | in vitro assessment of the gene expression of ezh 2 and p300 during motor neuron differentiation of human umbilical cord blood mesenchymal stem cells |
| topic | cord blood motor neuron p300 ezh-2 |
| url | http://bcn.iums.ac.ir/article-1-1984-en.html |
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