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Promoting effect of small molecules in cardiomyogenic and neurogenic differentiation of rat bone marrow-derived mesenchymal stem cells
Small molecules, growth factors, and cytokines have been used to induce differentiation of stem cells into different lineages. Similarly, demethylating agents can trigger differentiation in adult stem cells. Here, we investigated the in vitro differentiation of rat bone marrow mesenchymal stem cells...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Dove Medical Press
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4699543/ https://www.ncbi.nlm.nih.gov/pubmed/26766903 http://dx.doi.org/10.2147/DDDT.S89658 |
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author | Khanabdali, Ramin Saadat, Anbarieh Fazilah, Maizatul Bazli, Khairul Fidaa’ Khairul Qazi, Rida-e-Maria Khalid, Ramla Sana Hasan Adli, Durriyyah Sharifah Moghadamtousi, Soheil Zorofchian Naeem, Nadia Khan, Irfan Salim, Asmat Shamsuddin, ShamsulAzlin Ahmad Mohan, Gokula |
author_facet | Khanabdali, Ramin Saadat, Anbarieh Fazilah, Maizatul Bazli, Khairul Fidaa’ Khairul Qazi, Rida-e-Maria Khalid, Ramla Sana Hasan Adli, Durriyyah Sharifah Moghadamtousi, Soheil Zorofchian Naeem, Nadia Khan, Irfan Salim, Asmat Shamsuddin, ShamsulAzlin Ahmad Mohan, Gokula |
author_sort | Khanabdali, Ramin |
collection | PubMed |
description | Small molecules, growth factors, and cytokines have been used to induce differentiation of stem cells into different lineages. Similarly, demethylating agents can trigger differentiation in adult stem cells. Here, we investigated the in vitro differentiation of rat bone marrow mesenchymal stem cells (MSCs) into cardiomyocytes by a demethylating agent, zebularine, as well as neuronal-like cells by β-mercaptoethanol in a growth factor or cytokines-free media. Isolated bone marrow-derived MSCs cultured in Dulbecco’s Modified Eagle’s Medium exhibited a fibroblast-like morphology. These cells expressed positive markers for CD29, CD44, and CD117 and were negative for CD34 and CD45. After treatment with 1 μM zebularine for 24 hours, the MSCs formed myotube-like structures after 10 days in culture. Expression of cardiac-specific genes showed that treated MSCs expressed significantly higher levels of cardiac troponin-T, Nkx2.5, and GATA-4 compared with untreated cells. Immunocytochemical analysis showed that differentiated cells also expressed cardiac proteins, GATA-4, Nkx 2.5, and cardiac troponin-T. For neuronal differentiation, MSCs were treated with 1 and 10 mM β-mercaptoethanol overnight for 3 hours in complete and serum-free Dulbecco’s Modified Eagle’s Medium, respectively. Following overnight treatment, neuron-like cells with axonal and dendritic-like projections originating from the cell body toward the neighboring cells were observed in the culture. The mRNA expression of neuronal-specific markers, Map2, Nefl, Tau, and Nestin, was significantly higher, indicating that the treated cells differentiated into neuronal-like cells. Immunostaining showed that differentiated cells were positive for the neuronal markers Flk, Nef, Nestin, and β-tubulin. |
format | Online Article Text |
id | pubmed-4699543 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Dove Medical Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-46995432016-01-13 Promoting effect of small molecules in cardiomyogenic and neurogenic differentiation of rat bone marrow-derived mesenchymal stem cells Khanabdali, Ramin Saadat, Anbarieh Fazilah, Maizatul Bazli, Khairul Fidaa’ Khairul Qazi, Rida-e-Maria Khalid, Ramla Sana Hasan Adli, Durriyyah Sharifah Moghadamtousi, Soheil Zorofchian Naeem, Nadia Khan, Irfan Salim, Asmat Shamsuddin, ShamsulAzlin Ahmad Mohan, Gokula Drug Des Devel Ther Original Research Small molecules, growth factors, and cytokines have been used to induce differentiation of stem cells into different lineages. Similarly, demethylating agents can trigger differentiation in adult stem cells. Here, we investigated the in vitro differentiation of rat bone marrow mesenchymal stem cells (MSCs) into cardiomyocytes by a demethylating agent, zebularine, as well as neuronal-like cells by β-mercaptoethanol in a growth factor or cytokines-free media. Isolated bone marrow-derived MSCs cultured in Dulbecco’s Modified Eagle’s Medium exhibited a fibroblast-like morphology. These cells expressed positive markers for CD29, CD44, and CD117 and were negative for CD34 and CD45. After treatment with 1 μM zebularine for 24 hours, the MSCs formed myotube-like structures after 10 days in culture. Expression of cardiac-specific genes showed that treated MSCs expressed significantly higher levels of cardiac troponin-T, Nkx2.5, and GATA-4 compared with untreated cells. Immunocytochemical analysis showed that differentiated cells also expressed cardiac proteins, GATA-4, Nkx 2.5, and cardiac troponin-T. For neuronal differentiation, MSCs were treated with 1 and 10 mM β-mercaptoethanol overnight for 3 hours in complete and serum-free Dulbecco’s Modified Eagle’s Medium, respectively. Following overnight treatment, neuron-like cells with axonal and dendritic-like projections originating from the cell body toward the neighboring cells were observed in the culture. The mRNA expression of neuronal-specific markers, Map2, Nefl, Tau, and Nestin, was significantly higher, indicating that the treated cells differentiated into neuronal-like cells. Immunostaining showed that differentiated cells were positive for the neuronal markers Flk, Nef, Nestin, and β-tubulin. Dove Medical Press 2015-12-24 /pmc/articles/PMC4699543/ /pubmed/26766903 http://dx.doi.org/10.2147/DDDT.S89658 Text en © 2016 Khanabdali et al. This work is published by Dove Medical Press Limited, and licensed under Creative Commons Attribution – Non Commercial (unported, v3.0) License The full terms of the License are available at http://creativecommons.org/licenses/by-nc/3.0/. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. |
spellingShingle | Original Research Khanabdali, Ramin Saadat, Anbarieh Fazilah, Maizatul Bazli, Khairul Fidaa’ Khairul Qazi, Rida-e-Maria Khalid, Ramla Sana Hasan Adli, Durriyyah Sharifah Moghadamtousi, Soheil Zorofchian Naeem, Nadia Khan, Irfan Salim, Asmat Shamsuddin, ShamsulAzlin Ahmad Mohan, Gokula Promoting effect of small molecules in cardiomyogenic and neurogenic differentiation of rat bone marrow-derived mesenchymal stem cells |
title | Promoting effect of small molecules in cardiomyogenic and neurogenic differentiation of rat bone marrow-derived mesenchymal stem cells |
title_full | Promoting effect of small molecules in cardiomyogenic and neurogenic differentiation of rat bone marrow-derived mesenchymal stem cells |
title_fullStr | Promoting effect of small molecules in cardiomyogenic and neurogenic differentiation of rat bone marrow-derived mesenchymal stem cells |
title_full_unstemmed | Promoting effect of small molecules in cardiomyogenic and neurogenic differentiation of rat bone marrow-derived mesenchymal stem cells |
title_short | Promoting effect of small molecules in cardiomyogenic and neurogenic differentiation of rat bone marrow-derived mesenchymal stem cells |
title_sort | promoting effect of small molecules in cardiomyogenic and neurogenic differentiation of rat bone marrow-derived mesenchymal stem cells |
topic | Original Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4699543/ https://www.ncbi.nlm.nih.gov/pubmed/26766903 http://dx.doi.org/10.2147/DDDT.S89658 |
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