Iranian Biomedical Journal
مجله بیومدیکال ایران
IBJ
Basic Sciences
http://ibj.pasteur.ac.ir
1
admin
1028-852X
2008-823X
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10.61186/ibj
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8888
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en
jalali
1398
10
1
gregorian
2020
1
1
24
1
online
1
fulltext
en
Effect of Different Concentrations of Forskolin Along with Mature Granulosa Cell Co-Culturing on Mouse Embryonic Stem Cell Differentiation into Germ-Like Cells
Tissue Engineering and Cell Biology
Tissue Engineering and Cell Biology
مقاله کامل
Full Length/Original Article
<div style="text-align: justify;"><strong>Background:</strong> Germ cell development processes are influenced by soluble factors and intercellular signaling events between them and the neighboring somatic cells. More insight into the molecular biology of the germ cell development from embryonic stem (ES) cells and investigation of appropriate factors, specifically those targeting differentiation processes, is of great importance. In this study, we established an <em>in</em> <em>vitro</em> model with higher ES cell differentiation rate to germ cells, using adenylate cyclase activator, forskolin. <strong>Methods:</strong> ES cells were first cultured for five days, leading to embryoid body (EB) formation. Subsequently, the EB were dissociated and cultured for an additional three days in different forskolin concentrations of 5, 20, and 50 µM, with or without granulosa cells (GC) co-culture. On the 8<sup>th</sup> day, we analyzed the expressions of 5 germ cell-specific markers using quantitative real-time-PCR technique along with cell viability assay by MTT test. <strong>Results: </strong>Our results showed that in the GC-free cultures, a 50-µM concentration of forskolin resulted in a significant increase in <em>Mvh</em>, <em>Gdf9</em>, <em>Scp3</em>, and <em>Rec8</em> expression levels in comparison to the control. However, when the cells were co-cultured with the GCs, 20-µM concentration of forskolin could also increase the expression of those germ cell-specific marker genes. Furthermore, results from the MTT assay showed enhanced cell proliferation and survival at all three concentrations of forskolin, but 20-µM concentration was the most potent one. <strong>Conclusion:</strong> These data indicate that forskolin can stimulate differentiation and proliferation, dose-dependently; however, the influence of GCs <em>co-culturing </em>should not go unnoticed<em>.</em></div>
Embryonic stem cells, Forskolin, Germ cells, Granulosa cells
30
38
http://ibj.pasteur.ac.ir/browse.php?a_code=A-10-1-784&slc_lang=en&sid=1
Soghra
Bahmanpour
100319475328460054609
100319475328460054609
No
Laboratory for Stem Cell Research, Department of Anatomical Sciences, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
Amirhesam
Keshavarz
100319475328460054610
100319475328460054610
No
Laboratory for Stem Cell Research, Department of Anatomical Sciences, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
Nehleh
Zarei Fard
100319475328460054611
100319475328460054611
Yes
Laboratory for Stem Cell Research, Department of Anatomical Sciences, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran