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Reprogramming alters the gene expression in somatic cells, transforming them into induced pluripotent stem (iPS) cells over several generations. Scientists can reprogram cells by introducing genes for four transcription factors—Oct4, Sox2, Klf4, and c-Myc (OSKM) by viral or non-viral methods. These factors are also known as Yamanaka factors after Shinya Yamanaka, who first generated iPS cells using mouse skin cells. Yamanaka was awarded the Nobel Prize in Physiology or Medicine in 2012...
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Chromatin modification alters gene expression; therefore, scientists can add histone-modifying enzymes, histone variants, and chromatin remodeling complexes to somatic cells to aid reprogramming into pluripotent stem (iPS) cells.
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Nuclear reprogramming is a process of transforming one cell type into an unrelated cell type by epigenetic changes that alter the cell’s original gene expression pattern. Such epigenetic changes force cells to express a different set of genes, which play a significant role in inducing transformation into other cell types. Nuclear reprogramming offers applications in reproductive cloning for livestock propagation and regenerative medicine — developing patient-specific cells for...
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The cells of the blastocyst inner cell mass only remain pluripotent for a short time. This state of pluripotency and self-renewal can be maintained in embryonic stem (ES) cell culture by adding specific chemicals or growth factors to ensure the cells can continue dividing and later differentiate into different cell types. In some cases, the cells are grown on a feeder layer of differentiated cells, which provides the growth factors and extracellular matrix components necessary for stem cell...
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Stem cells are undifferentiated cells that divide and produce different types of cells. Ordinarily, cells that have differentiated into a specific cell type are post-mitotic—that is, they no longer divide. However, scientists have found a way to reprogram these mature cells so that they “de-differentiate” and return to an unspecialized, proliferative state. These cells are also pluripotent like embryonic stem cells—able to produce all cell types—and are therefore...
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Manipulating the Mediator complex to induce naïve pluripotency.

Cian J Lynch1, Raquel Bernad1, Isabel Calvo1

  • 1Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology (BIST), Barcelona, Spain.

Experimental Cell Research
|August 11, 2020
PubMed
Summary

Researchers stabilized human naïve pluripotent stem cells (PSCs) using a novel chemical inhibitor. This method enhances cellular identity and developmental potential, overcoming challenges in PSC maintenance.

Keywords:
2iCDK8DemethylationEmbryonic stemMEKMediatorNaïvePhase separationPluripotencyPrimedRNA polymeraseReprogrammingSuper-enhancers

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Chemical Reversion of Conventional Human Pluripotent Stem Cells to a Naïve-like State with Improved Multilineage Differentiation Potency
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Area of Science:

  • Stem Cell Biology
  • Molecular and Cellular Biology

Background:

  • Human pluripotent stem cells (PSCs) exist in primed or naïve states, with naïve PSCs offering a more homogenous and optimal starting point for research.
  • Maintaining and expanding human naïve PSCs is challenging due to their inherent instability and heterogeneity compared to primed PSCs.

Purpose of the Study:

  • To develop a strategy for stabilizing human PSCs in the naïve state.
  • To investigate the efficacy of inhibiting Cyclin-Dependent Kinases 8 and 19 (CDK8/19) for naïve PSC stabilization.

Main Methods:

  • Utilized a specific chemical inhibitor targeting CDK8 and CDK19 (CDK8/19i).
  • Assessed the impact of CDK8/19 inhibition on enhancer activity and transcriptional programs in human PSCs.
  • Evaluated chromosomal stability and developmental potential of CDK8/19i-stabilized naïve human PSCs during long-term expansion.

Main Results:

  • CDK8/19 inhibition globally stimulates enhancers, reinforcing transcriptional programs essential for cellular identity.
  • CDK8/19i treatment efficiently stabilizes human PSCs in the naïve state.
  • Unlike previous methods, CDK8/19i-naïve PSCs exhibit chromosomal stability and retain developmental potential after prolonged culture, potentially due to the absence of DNA demethylation.

Conclusions:

  • CDK8/19 inhibition provides a robust method for stabilizing human PSCs in the naïve pluripotent state.
  • This approach overcomes key limitations of previous methods, offering chromosomally stable naïve PSCs with retained developmental potential.
  • The findings suggest that CDK8/19 inhibition could be a valuable tool for stem cell research and therapeutic applications, with potential implications for other cell fate decisions.