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Cell Journal، جلد ۱۶، شماره Suppl ۱، صفحات ۱۵۷-۱۵۷
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عنوان انگلیسی Ps-126: The Role of Sonic Hedgehog on Differentiation of Definitive Endoderm from Mouse Embryonic Stem Cells
چکیده انگلیسی مقاله Objective: Sonic hedgehog (Shh) affects the development of embryo and also embryonic stem cells (ESCs). The aim of this study was to determine the effect of Shh inhibition during definitive endoderm differentiation of mouse embryonic stem cells. Materials and Methods: The mouse embryonic stem cell line (MUKF3) was cultured for 5 days in non adherent petri dishes to form embryoid bodies (EBs). The media of control group (group I) contained DMEM/ F12+%10 knockout serum replacement (KOSR). Experimental group (group II) received the hedgehog interacting protein (HIP) (10 μg/ml). Then EBs were cultured on gelatin coated dishes in attached position for further 8 days. Total RNA of each group was extracted and the expression of target genes was compared between groups by relative RT-PCR method and statistically analyzed. Also immunocytochemistry staining of Goosecoid and Sox-17 of treated EBs was done to identify the definitive endoderm cells. Results: There was a significant decrease in expression of oct-4, nanog, brachury, shh, gata4 and goosecoid in the experimental group (p< 0.005), while the expression of pdx-1 and tat in experimental group were significantly increased (p< 0.005). Immunocytochemistry revealed that in the experimental group the amount of Goosecoid was lower in comparison to the control group but it seems that there was no difference for Sox- 17. Conclusion: The study presents Shh inhibition as a molecular pathway for induction of definitive endoderm from ESCs by affecting the fate of mesoendodermal cells.
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/451
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