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Related Concept Videos

Methods of Nuclear Reprogramming01:24

Methods of Nuclear Reprogramming

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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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Introduction to Nuclear Reprogramming01:14

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Nuclear reprogramming is the process of switching gene expression of one cell type to that of another cell type, usually from a differentiated cell state to an undifferentiated cell state. Differentiation occurs during processes such as development and morphogenesis, tissue regeneration, and malignancy. Cells can also be artificially induced to reprogram their gene expression by techniques such as nuclear transfer, induced pluripotency, and cell fusion. Such techniques have many applications in...
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Somatic to iPS Cell Reprogramming01:29

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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 in iPS Cells01:32

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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.
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...
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Kinetic Measurement and Real Time Visualization of Somatic Reprogramming
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Tracking and Predicting Human Somatic Cell Reprogramming Using Nuclear Characteristics.

Kaivalya Molugu1, Ty Harkness2, Jared Carlson-Stevermer2

  • 1Graduate Program in Biophysics, University of Wisconsin-Madison, Madison, Wisconsin; Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, Wisconsin; Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, Wisconsin.

Biophysical Journal
|November 9, 2019
PubMed
Summary
This summary is machine-generated.

Researchers developed a micropatterned substrate to track human cell reprogramming in real-time. This method aids in identifying and isolating fully reprogrammed induced pluripotent stem cells (iPSCs) for therapeutic applications.

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Area of Science:

  • Stem cell biology
  • Biotechnology
  • Cellular reprogramming

Background:

  • Induced pluripotent stem cells (iPSCs) are crucial for disease modeling, toxicology, cell therapy, and regenerative medicine.
  • Current methods for iPSC identification and enrichment often disrupt the cellular microenvironment.
  • The reprogramming process is inherently stochastic and inefficient, yielding heterogeneous cell populations.

Purpose of the Study:

  • To develop a novel method for real-time monitoring of human cell reprogramming.
  • To preserve the cellular microenvironment during reprogramming analysis.
  • To identify predictive markers for distinguishing fully reprogrammed iPSCs from intermediates.

Main Methods:

  • Development of a micropatterned substrate for live-cell microscopy.
  • Confining cells into discrete islands to maintain microenvironment.
  • High-content analysis to identify nuclear characteristics.
  • Creation of a computational model to predict reprogramming progression.

Main Results:

  • The micropatterned substrate enabled dynamic live-cell microscopy of reprogramming.
  • A combination of eight nuclear characteristics accurately predicted reprogramming status.
  • The model successfully distinguished partially reprogrammed cells from fully reprogrammed iPSCs.
  • Preservation of cellular microenvironment cues was achieved.

Conclusions:

  • Micropatterned substrates offer a powerful tool for in situ tracking of cellular reprogramming.
  • This approach can enhance the biomanufacturing of therapeutically relevant iPSCs.
  • The method facilitates the study of multiscale cellular changes during human cell fate transitions.