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Related Concept Videos

Maintenance of the ES Cell State01:14

Maintenance of the ES Cell State

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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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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Human Pluripotent Stem Cell Culture on Polyvinyl Alcohol-Co-Itaconic Acid Hydrogels with Varying Stiffness Under Xeno-Free Conditions
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Substrate stiffness affects early differentiation events in embryonic stem cells.

Nicholas D Evans1, Caterina Minelli, Eileen Gentleman

  • 1Department of Materials, Imperial College London, London, UK.

European Cells & Materials
|September 22, 2009
PubMed
Summary

Substrate stiffness influences embryonic stem cell differentiation. Stiffer environments promote cell growth, gene expression, and osteogenic differentiation, highlighting the role of mechanosensing in development.

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Area of Science:

  • Developmental Biology
  • Stem Cell Biology
  • Biomaterials Science

Background:

  • Embryonic stem cells (ESCs) are crucial for regenerative medicine and studying early development.
  • Cell differentiation is influenced by chemical cues and mechanical properties of the microenvironment.
  • Mechanosensing, the ability of cells to detect and respond to mechanical stimuli, is increasingly recognized in cellular processes.

Purpose of the Study:

  • To investigate the hypothesis that substrate stiffness influences embryonic stem cell (ESC) differentiation.
  • To determine the effects of varying substrate stiffness on ESC attachment, spreading, growth, and differentiation.
  • To explore the role of the mechanical environment in both early and terminal ESC differentiation.

Main Methods:

  • Culturing murine ESCs on polydimethylsiloxane (PDMS) substrates with controlled, varying stiffness.
  • Assessing cell attachment, spreading, and growth rates in response to substrate stiffness.
  • Quantifying the expression of key developmental genes (Brachyury, Mixl1, Eomes) via gene expression analysis.
  • Evaluating osteogenic differentiation potential on substrates of different stiffness.

Main Results:

  • Cell attachment was independent of substrate stiffness.
  • Cell spreading and growth increased significantly with increasing substrate stiffness.
  • Genes associated with primitive streak formation and mesendoderm differentiation (Brachyury, Mixl1, Eomes) were upregulated on stiffer substrates.
  • Osteogenic differentiation of ESCs was enhanced on stiff substrates compared to soft substrates.

Conclusions:

  • Substrate stiffness plays a significant role in regulating ESC differentiation, affecting early developmental gene expression and terminal differentiation.
  • Cellular mechanosensing is fundamental to mammalian development.
  • Mechanical properties of the environment must be considered in tissue engineering and in vitro cell culture for therapeutic applications.