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Kinetics of Gene Expression During Exposure of Mouse Stem Cells to Activin A

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Author(s): Mahmoud Hashemi-Tabar | Mahmoud Orazizadeh | Ali Ghanbari | Fereshteh Negad Dehbashi

Journal: Pakistan Journal of Biological Sciences
ISSN 1028-8880

Volume: 12;
Issue: 4;
Start page: 324;
Date: 2009;
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Keywords: Activin A | definitive endoderm | nanog | sonic hedgehog | stem cells

ABSTRACT
This study aimed to evaluate the pattern of gene expression induced by activin A in mouse Embryonic Stem Cells (ESCs). Mouse ES cells cultured in undifferentiated state by leukemia inhibitory factor and feeder layer cells. Following removing these two anti differentiation factors for 5 days and forming Embryoid Bodies (EBs), the cells divided to 8 equal cells per groups. Differentiation procedure was performed in a two staged protocol; Formed EBs for 4 days (Stage one); expanded differentiated ESCs on gelatin coated dishes for one week (stage two). In the stage one, the media of groups 2-7 contained 10, 30 and 100 ng mL-1 Activin A. The media in stage two was the same for all groups and contained only Fetal Bovine Serum (FBS). The expression of undifferentiated, ectoderm, mesoderm and endoderm markers were compared with relative RT-PCR method and statistically analyzed. The expression of an undifferentiating marker; Nanog was increased in the Activin A treated groups of stage one. The expression of OCT4 reduced in Activin A treated groups in stage two. In the stage one, the expression of Nodal increased by Activin A. expression of sonic hedgehog (Shh) was suppressed in Activin A treated groups of both stages. In stage two, there were significant decrease for the expression of mesoderm (Brachyury) and Nodal and visceral endoderm (GATA4) markers (p
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