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Embryonic Stem Cells00:58

Embryonic Stem Cells

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Embryonic stem (ES) cells are undifferentiated pluripotent cells, meaning they can produce any cell type in the body. This gives them tremendous potential in science and medicine since they can generate specific cell types for use in research or to replace body cells lost due to damage or disease.
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Embryonic Stem Cells00:57

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Embryonic stem (ES) cells were first discovered in mice in 1981 by Martin Evans. In 1998, James Thomson identified a method to isolate embryonic stem cells from humans. Human embryonic stem cells (hESCs) are obtained from 3-5 day old embryos that remain unused after an in vitro fertilization procedure.
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Multicellular organisms contain a variety of structurally and functionally distinct cell types, but the DNA in all the cells originated from the same parent cells. The differences in the cells can be attributed to the differential gene expression. Liver cells, whose functions include detoxification of blood, production of bile to metabolize fats, and synthesis of proteins essential for metabolism, must express a specific set of genes to perform their functions. Gene expression also varies with...
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Gene expression is the process in which DNA directs the synthesis of functional products, that is, proteins. Cells can regulate gene expression at various stages. It allows organisms to generate different cell types and enables cells to adapt to internal and external factors.
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A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is comprised  of nucleotides and proteins are comprised of amino acids, a mediator is required to convert the information encoded in DNA into proteins. This mediator is the messenger RNA (mRNA). mRNA copies the blueprint from DNA by a process called transcription. In eukaryotes, transcription occurs in the nucleus by complementary base-pairing with the DNA template. The mRNA is then...
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Updated: Feb 9, 2026

Derivation of Hematopoietic Stem Cells from Murine Embryonic Stem Cells
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Temporal Gene Expression and DNA Methylation during Embryonic Stem Cell Derivation.

Azam Samadian1, Mahdi Hesaraki1, Sepideh Mollamohammadi1

  • 1Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.

Cell Journal
|May 31, 2018
PubMed
Summary

Dual inhibition of MEK and TGF-β signaling pathways (R2i) efficiently generates mouse embryonic stem cells (ESCs). This method promotes pluripotency gene expression and reduces differentiation gene expression by altering DNA methylation patterns during derivation.

Keywords:
DNA MethylationMEK InhibitorMouse Embryonic Stem CellsR2iTGFβ Inhibitor

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Area of Science:

  • Stem cell biology
  • Developmental biology
  • Epigenetics

Background:

  • Embryonic stem cell (ESC) generation is crucial for regenerative medicine and developmental studies.
  • Current methods often involve serum, which can lead to differentiation.
  • Dual inhibition of MEK and TGF-β signaling offers a promising alternative for efficient ESC derivation.

Purpose of the Study:

  • To investigate the molecular mechanisms underlying mouse ESC derivation using dual inhibition of MEK and TGF-β signaling (R2i).
  • To compare gene expression and DNA methylation patterns between R2i and conventional serum conditions.

Main Methods:

  • Zona-free E3.5 blastocysts were cultured on MEF feeders in R2i or serum media.
  • Inner cell mass (ICM) and ESCs were collected at different time points.
  • Quantitative real-time PCR (qRT-PCR) and DNA methylation analysis were performed.

Main Results:

  • R2i treatment significantly upregulated pluripotency genes (Oct4, Nanog, Sox2) and downregulated differentiation genes (Gata6, Cdx2).
  • DNA methylation analysis revealed dynamic changes in Oct4 and Nanog promoter regions.
  • Lower DNA methylation in R2i-derived ESCs correlated with higher pluripotency gene expression compared to serum conditions.

Conclusions:

  • Inhibiting MEK and TGF-β signaling pathways during the initial 5 days of derivation promotes ground-state pluripotency in ESCs.
  • R2i conditions facilitate ESC generation by modulating gene expression through epigenetic modifications.
  • This approach offers a more efficient and controlled method for generating high-quality ESCs.