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Somatic to iPS Cell Reprogramming01:29

Somatic to iPS Cell Reprogramming

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 for this...
Methods of Nuclear Reprogramming01:24

Methods of Nuclear Reprogramming

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 injury repair.
Chromatin Modification in iPS Cells01:32

Chromatin Modification in iPS Cells

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.
Compact chromatin makes reprogramming difficult. Enzymes, such as histone demethylases and acetyltransferases, are often added during reprogramming to loosen the chromatin, making the DNA more accessible to transcription factors. Molecules that inhibit histone...
iPS Cell Differentiation01:22

iPS Cell Differentiation

The ability of induced pluripotent stem cells or iPSCs to differentiate into most body cell types has stimulated repair and regenerative medicine research over the past few decades. iPSC-derived blood cells, hepatocytes, beta islet cells, cardiomyocytes, neurons, and other cell types can repair injuries or regenerate damaged tissue in diseases such as diabetes and neurodegenerative disorders.
Induced Pluripotent Stem Cells01:06

Induced Pluripotent Stem Cells

Stem cells are undifferentiated cells that divide and produce different cell types. Ordinarily, cells that have differentiated into a specific cell type are terminally differentiated; however, scientists have found a way to reprogram these mature cells so that they dedifferentiate and return to an unspecialized, proliferative state. These cells are pluripotent like embryonic stem cells—able to produce all cell types—and are called induced pluripotent stem cells (iPSCs).
Somatic cells are...
Induced Pluripotent Stem Cells01:13

Induced Pluripotent Stem Cells

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 called induced pluripotent stem...

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Related Experiment Video

Updated: Jun 21, 2026

Cell Surface Marker Mediated Purification of iPS Cell Intermediates from a Reprogrammable Mouse Model
10:32

Cell Surface Marker Mediated Purification of iPS Cell Intermediates from a Reprogrammable Mouse Model

Published on: September 6, 2014

The Ink4/Arf locus is a barrier for iPS cell reprogramming.

Han Li1, Manuel Collado, Aranzazu Villasante

  • 1Tumor Suppression Group, Spanish National Cancer Research Centre (CNIO), 3 Melchor Fernandez Almagro Street, Madrid E-28029, Spain.

Nature
|August 12, 2009
PubMed
Summary

Silencing the Ink4/Arf locus is crucial for induced pluripotent stem (iPS) cell generation. Transient inhibition of this locus significantly improves reprogramming efficiency and kinetics, offering a key strategy for iPS cell production.

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Reprogramming Human Somatic Cells into Induced Pluripotent Stem Cells (iPSCs) Using Retroviral Vector with GFP
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Reprogramming Human Somatic Cells into Induced Pluripotent Stem Cells (iPSCs) Using Retroviral Vector with GFP

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Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor
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Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor

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Last Updated: Jun 21, 2026

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Published on: September 6, 2014

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08:25

Reprogramming Human Somatic Cells into Induced Pluripotent Stem Cells (iPSCs) Using Retroviral Vector with GFP

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Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor
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Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor

Published on: February 1, 2015

Area of Science:

  • Cellular reprogramming
  • Epigenetics
  • Stem cell biology

Background:

  • Induced pluripotent stem (iPS) cell generation mechanisms involving Oct4, Klf4, and Sox2 are not fully understood.
  • The Ink4/Arf locus, encoding tumor suppressors p16(Ink4a), p19(Arf), and p15(Ink4b), is upregulated in differentiated cells.
  • This locus's role in reprogramming efficiency is a critical area of investigation.

Purpose of the Study:

  • To investigate the role of the Ink4/Arf locus in the reprogramming of differentiated cells into iPS cells.
  • To determine if silencing the Ink4/Arf locus is essential for efficient iPS cell generation.
  • To explore therapeutic strategies for enhancing iPS cell production by targeting the Ink4/Arf locus.

Main Methods:

  • Analysis of Ink4/Arf locus silencing in iPS cells and embryonic stem (ES) cells.
  • Assessment of Ink4/Arf locus expression during reprogramming under various cell culture conditions.
  • Genetic and shRNA-mediated inhibition of the Ink4/Arf locus to evaluate reprogramming efficiency.
  • Comparative analysis of Ink4/Arf locus function in murine and human cells.
  • Investigation of Ink4/Arf locus regulation in aged cells and its impact on reprogramming.

Main Results:

  • The Ink4/Arf locus is completely silenced in iPS and ES cells, acquiring bivalent chromatin marks.
  • Reprogramming conditions enhance Ink4/Arf expression, necessitating its silencing for efficient iPS cell generation.
  • Cooperative action of Oct4, Klf4, and Sox2 represses the Ink4/Arf locus, correlating with stemness markers.
  • Genetic inhibition of Ink4/Arf significantly boosts iPS cell generation kinetics and colony numbers.
  • Arf is the primary barrier in murine cells (via p53/p21), while INK4a is more critical in human fibroblasts.
  • Age-related upregulation of Ink4/Arf reduces reprogramming efficiency, which is rescued by locus inhibition.

Conclusions:

  • Ink4/Arf locus silencing is a rate-limiting step in iPS cell reprogramming.
  • Transient inhibition of the Ink4/Arf locus is a promising strategy to improve iPS cell generation.
  • Understanding Ink4/Arf regulation provides insights into cellular plasticity and aging.
  • Targeting Ink4/Arf offers a potential therapeutic avenue for regenerative medicine applications.