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RNA-based Reprogramming of Human Primary Fibroblasts into Induced Pluripotent Stem Cells
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Actin stress in cell reprogramming.

Jun Guo1, Yuexiu Wang2, Frederick Sachs3

  • 1Department of Biochemistry, Nanjing Medical University, Nanjing, Jiangsu 210029, China;

Proceedings of the National Academy of Sciences of the United States of America
|November 26, 2014
PubMed
Summary
This summary is machine-generated.

Cell mechanics influences stem cell reprogramming. A new probe, AcpA, reveals increased actin force during stemness, enabling real-time cell force measurement and understanding of cellular processes.

Keywords:
actincell mechanicsforce probereprogrammingstem cell

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Area of Science:

  • Cell Biology
  • Biophysics
  • Stem Cell Biology

Background:

  • Cell mechanics is crucial for stem cell reprogramming and differentiation.
  • Understanding forces within actin, a key structural protein, is vital for cell physiology.

Purpose of the Study:

  • To develop a genetically encoded optical probe (AcpA) for real-time measurement of forces in cellular actin.
  • To investigate the relationship between actin forces and stem cell states.

Main Methods:

  • Creation of actin-cpstFRET-actin (AcpA), a FRET sensor flanked by g-actin subunits.
  • Utilizing AcpA to measure actin forces in various cell types (HEK-293, MDCK) under different substrate stiffness conditions.
  • Assessing cell viability and actin distribution of labeled actin.

Main Results:

  • Stemness in cells correlated with increased actin force.
  • Softening the substrate induced stem-like cell reprogramming in HEK-293 cells.
  • MDCK cell reprogramming required a soft substrate and specific gene expression.
  • Replating cells on stiff substrates led to redifferentiation and reduced actin force.
  • Actinin force measurements indicated protein-specific force dynamics.

Conclusions:

  • The AcpA probe is a viable tool for measuring actin forces in living cells without affecting cell physiology.
  • Actin force is a key mechanical regulator in stem cell reprogramming and differentiation.
  • Protein-specific force measurements are necessary for a comprehensive understanding of cell mechanics.