Differentiation of endometrial stem cells into motor neurons by the use of purmorphamin small molecule
Background: Small molecule Purmorphamin (PMA) is the agonist of smoothened protein in Sonic hedgehog (Shh) signaling pathway. Effect of purmorphamin small molecule on differentiation of mesenchymal cells into bone tissue has been studied previously. Use of Shh causes progression of neural differenti...
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doaj-36cd1aa721de44419c94d16138d6650e2020-11-24T23:53:49ZfasTehran University of Medical SciencesTehran University Medical Journal1683-17641735-73222017-03-017412861868Differentiation of endometrial stem cells into motor neurons by the use of purmorphamin small moleculeHoma Mohseni Kouchesfahani0Somayeh Ebrahimi-Barough1Jafar Ai2Azam Rahimi3 Department of Animal Biology, School of Biological Sciences, Kharazmi University, Tehran, Iran. Department of Tissue Engineering and Applied Cell Sciences, School of Advance Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran. Department of Tissue Engineering and Applied Cell Sciences, School of Advance Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran. Department of Animal Biology, Cell & Developmental Biology, School of Biological Sciences, Kharazmi University, Tehran, Iran. Background: Small molecule Purmorphamin (PMA) is the agonist of smoothened protein in Sonic hedgehog (Shh) signaling pathway. Effect of purmorphamin small molecule on differentiation of mesenchymal cells into bone tissue has been studied previously. Use of Shh causes progression of neural differentiation, and the differentiated cells express specific neural markers. Neurofilament (NF) and acetylcholine esterase (Chat) are specific markers of motor neurons and their expression in differentiated cells indicates their conversion into motor neurons. The aim of this study was to evaluate the ability of PMA to differentiate the human endometrial stem cells (hEnSCs) into motor neurons. Methods: This analytical study was done in Tehran University of Medical Sciences laboratory on September of 2015. In this study hEnSCs were enzymatically extracted from endometrial tissue. After third passages, the flow cytometry was done for mesenchymal stem cells markers. The mesenchymal stem cells were divided into control and differentiated groups. FBS 10%+DMEM/F12 was added to the culture medium of control group and the differentiating group was treated with differentiating medium containing N2, PMA, DMEM/F12, FBS, B27, IBMX, 2ME, FGF2, RA, BDNF. After 21 days immunocytochemistry (ICC) test was done for the expression of NF and Chat proteins and Real-time PCR analysis for expression of neural markers such as NF, Chat, Nestin and GFAP (as glial marker) at mRNA level. Results: The flow cytometry analysis showed that hEnSCs were positive for mesenchymal markers CD90, CD105 and CD146 and negative for endothelial marker CD31, and hematopoietic marker CD34. The immunocytochemistry and Real time-PCR results showed that the cells treated with PMA expressed motor neuron markers of NF and Chat. Conclusion: According to the results of this study, it can be concluded that small molecule PMA has the potency to induce the differentiation of hEnSCs into neural cells, specifically motor neurons by activating Shh signaling pathway.http://tumj.tums.ac.ir/browse.php?a_code=A-10-25-5544&slc_lang=en&sid=1differentiation endometrial stem cells purmorphamin sonic hedgehog pathway |
collection |
DOAJ |
language |
fas |
format |
Article |
sources |
DOAJ |
author |
Homa Mohseni Kouchesfahani Somayeh Ebrahimi-Barough Jafar Ai Azam Rahimi |
spellingShingle |
Homa Mohseni Kouchesfahani Somayeh Ebrahimi-Barough Jafar Ai Azam Rahimi Differentiation of endometrial stem cells into motor neurons by the use of purmorphamin small molecule Tehran University Medical Journal differentiation endometrial stem cells purmorphamin sonic hedgehog pathway |
author_facet |
Homa Mohseni Kouchesfahani Somayeh Ebrahimi-Barough Jafar Ai Azam Rahimi |
author_sort |
Homa Mohseni Kouchesfahani |
title |
Differentiation of endometrial stem cells into motor neurons by the use of purmorphamin small molecule |
title_short |
Differentiation of endometrial stem cells into motor neurons by the use of purmorphamin small molecule |
title_full |
Differentiation of endometrial stem cells into motor neurons by the use of purmorphamin small molecule |
title_fullStr |
Differentiation of endometrial stem cells into motor neurons by the use of purmorphamin small molecule |
title_full_unstemmed |
Differentiation of endometrial stem cells into motor neurons by the use of purmorphamin small molecule |
title_sort |
differentiation of endometrial stem cells into motor neurons by the use of purmorphamin small molecule |
publisher |
Tehran University of Medical Sciences |
series |
Tehran University Medical Journal |
issn |
1683-1764 1735-7322 |
publishDate |
2017-03-01 |
description |
Background: Small molecule Purmorphamin (PMA) is the agonist of smoothened protein in Sonic hedgehog (Shh) signaling pathway. Effect of purmorphamin small molecule on differentiation of mesenchymal cells into bone tissue has been studied previously. Use of Shh causes progression of neural differentiation, and the differentiated cells express specific neural markers. Neurofilament (NF) and acetylcholine esterase (Chat) are specific markers of motor neurons and their expression in differentiated cells indicates their conversion into motor neurons. The aim of this study was to evaluate the ability of PMA to differentiate the human endometrial stem cells (hEnSCs) into motor neurons.
Methods: This analytical study was done in Tehran University of Medical Sciences laboratory on September of 2015. In this study hEnSCs were enzymatically extracted from endometrial tissue. After third passages, the flow cytometry was done for mesenchymal stem cells markers. The mesenchymal stem cells were divided into control and differentiated groups. FBS 10%+DMEM/F12 was added to the culture medium of control group and the differentiating group was treated with differentiating medium containing N2, PMA, DMEM/F12, FBS, B27, IBMX, 2ME, FGF2, RA, BDNF. After 21 days immunocytochemistry (ICC) test was done for the expression of NF and Chat proteins and Real-time PCR analysis for expression of neural markers such as NF, Chat, Nestin and GFAP (as glial marker) at mRNA level.
Results: The flow cytometry analysis showed that hEnSCs were positive for mesenchymal markers CD90, CD105 and CD146 and negative for endothelial marker CD31, and hematopoietic marker CD34. The immunocytochemistry and Real time-PCR results showed that the cells treated with PMA expressed motor neuron markers of NF and Chat.
Conclusion: According to the results of this study, it can be concluded that small molecule PMA has the potency to induce the differentiation of hEnSCs into neural cells, specifically motor neurons by activating Shh signaling pathway. |
topic |
differentiation endometrial stem cells purmorphamin sonic hedgehog pathway |
url |
http://tumj.tums.ac.ir/browse.php?a_code=A-10-25-5544&slc_lang=en&sid=1 |
work_keys_str_mv |
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