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Information Journal Paper

Title

TEMPORAL GENE EXPRESSION AND DNA METHYLATION DURING EMBRYONIC STEM CELL DERIVATION

Pages

  361-368

Abstract

 Objective: Dual inhibition of mitogen-activated protein kinase (MAPK) kinase (also known as MEK) and transforming growth factor b (TGFb) type I receptors by PD0325901 and SB431542, known as R2I has been introduced as a highly efficient approach to the generation of MOUSE EMBRYONIC STEM CELLS (ESC). In the present study, we investigated the molecular mechanisms underlying ESC derivation in the R2I condition.Materials and Methods: In this experimental study, zona-free whole E3.5 blastocysts were seeded on mouse embryonic fibroblast (MEF) feeder cells in both R2I and serum conventional media. The isolated inner cell mass (ICM), ESCs and the ICM-outgrowths were collected on days 3, 5 and 7 post-blastocyst culture for quantitative real timepolymerase chain reaction (qRT-PCR) analysis as well as to assess the DNA METHYLATION status at the time points during the transition from ICM to ESC.Results: qRT-PCR revealed a significantly higher expression of the pluripotency-related genes (Oct4, Nanog, Sox2, Rex1, Dppa3, Tcf3, Utf1, Nodal, Dax1, Sall4and b-Catenin) and lower expression of early differentiation genes (Gata6, Lefty2and Cdx2) in R2I condition compared to the serum condition. Moreover, the upstream region of Oct4 and Nanog showed a progressive increase in methylation levels in the upstream regions of the genes following in R2I or serum conditions, followed by a decrease of DNA METHYLATION in ESCs obtained under R2I. However, the methylation level of ICM outgrowths in the serum condition was much higher than R2I, at levels that could have a repressive effect and therefore explain the absence of expression of these two genes in the serum condition.Conclusion: Our investigation revealed that generation of ESCs in the ground-state of pluripotency could be achieved by inhibiting the MEK and TGF-b signaling pathways in the first 5 days of ESC derivation.

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    APA: Copy

    SAMADIAN, AZAM, HESARAKI, MAHDI, Mollamohammadi, Sepideh, Asgarian, Behrouz, TOTONCHI, MEHDI, & BAHARVAND, HOSSEIN. (2018). TEMPORAL GENE EXPRESSION AND DNA METHYLATION DURING EMBRYONIC STEM CELL DERIVATION. CELL JOURNAL (YAKHTEH), 20(3), 361-368. SID. https://sid.ir/paper/715023/en

    Vancouver: Copy

    SAMADIAN AZAM, HESARAKI MAHDI, Mollamohammadi Sepideh, Asgarian Behrouz, TOTONCHI MEHDI, BAHARVAND HOSSEIN. TEMPORAL GENE EXPRESSION AND DNA METHYLATION DURING EMBRYONIC STEM CELL DERIVATION. CELL JOURNAL (YAKHTEH)[Internet]. 2018;20(3):361-368. Available from: https://sid.ir/paper/715023/en

    IEEE: Copy

    AZAM SAMADIAN, MAHDI HESARAKI, Sepideh Mollamohammadi, Behrouz Asgarian, MEHDI TOTONCHI, and HOSSEIN BAHARVAND, “TEMPORAL GENE EXPRESSION AND DNA METHYLATION DURING EMBRYONIC STEM CELL DERIVATION,” CELL JOURNAL (YAKHTEH), vol. 20, no. 3, pp. 361–368, 2018, [Online]. Available: https://sid.ir/paper/715023/en

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