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Induction and Analysis of Epithelial to Mesenchymal Transition
Published on: August 27, 2013
Epithelial-mesenchymal transition events during human embryonic stem cell differentiation
Angela M Eastham1, Helen Spencer, Francesca Soncin
1Centre for Molecular Medicine, Faculty of Medical and Human Sciences, The University of Manchester, M13 9PT, United Kingdom.
Cancer Research
|December 7, 2007
Summary
Human embryonic stem cells (ES cells) exhibit epithelial-mesenchymal transition (EMT) characteristics during differentiation, involving E-cadherin changes and increased cell motility. Blocking E-cadherin in undifferentiated ES cells mimics EMT without inducing differentiation.
Area of Science:
- Cell Biology
- Developmental Biology
- Cancer Research
Background:
- Epithelial-mesenchymal transition (EMT) is crucial for embryonic development and cancer metastasis.
- During EMT, E-cadherin is downregulated, increasing cell motility and invasion.
- Human embryonic stem cells (ES cells) offer a model to study developmental processes.
Purpose of the Study:
- To investigate EMT-like events during human ES cell differentiation.
- To explore the role of E-cadherin in maintaining the undifferentiated state of human ES cells.
- To utilize human ES cells as a model for studying EMT in development and metastasis.
Main Methods:
- Monolayer culture of human ES cells.
- Analysis of cell differentiation markers (E-cadherin, N-cadherin, vimentin, Snail, Slug, OCT-4).
- Use of neutralizing antibody (nAb) SHE78.7 to block E-cadherin function.
- Assessment of cell motility, morphology, and antigen presentation (5T4).
Main Results:
- Human ES cell differentiation involves an E- to N-cadherin switch, increased vimentin, Snail/Slug, and matrix metalloproteinase activity, alongside enhanced motility.
- Blocking E-cadherin in undifferentiated ES cells induced motility, mesenchymal phenotype, and 5T4 antigen expression without affecting OCT-4 or EMT transcripts.
- Removal of the E-cadherin blocking antibody restored cell-cell contact, suppressed 5T4, and allowed differentiation into three germ layers.
Conclusions:
- E-cadherin stabilizes the actin cytoskeleton in human ES cells, preventing 5T4 cell surface localization.
- Human ES cells provide a valuable model for studying EMT relevant to both embryonic development and tumor metastasis.
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