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A High-throughput Cell Microarray Platform for Correlative Analysis of Cell Differentiation and Traction Forces
Published on: March 1, 2017
Dimeric integrin alpha5beta1 ligands confer morphological and differentiation responses to murine embryonic stem
Mark D Singh1, Michaela Kreiner, Clive S McKimmie
1Division of Immunology Infection and Inflammation, Glasgow Biomedical Research Centre, University of Glasgow, 120 University Place, Glasgow G12 8TA, UK.
Biochemical and Biophysical Research Communications
|October 17, 2009
Summary
Self-assembling ligands targeting integrin alpha5beta1 promote murine embryonic stem cell adhesion and pluripotency. Dimeric ligands enhanced cell colonies and influenced differentiation markers, offering insights for cell and tissue engineering.
Area of Science:
- Biomaterials Science
- Stem Cell Biology
- Cellular Engineering
Background:
- Murine embryonic stem cell (mES) adhesion and pluripotency are regulated by extracellular matrix interactions.
- Integrin alpha5beta1 is a key receptor mediating mES cell attachment to fibronectin.
- Controlling mES cell behavior requires precise presentation of integrin ligands.
Purpose of the Study:
- To investigate the functional relevance of self-assembling polyvalent ligands for integrin alpha5beta1 in mES cell adhesion.
- To generate clustered integrin alpha5beta1 ligand surfaces using fibronectin type III domains (FIII9'10).
- To assess the impact of these surfaces on mES cell morphology, pluripotency, and differentiation potential.
Main Methods:
- Synthesis of dimeric, trimeric, and tetrameric FIII9'10 ligands.
- Culture of mES cells on surfaces functionalized with FIII9'10 ligands and gelatin.
- Analysis of mES cell colony formation, morphology, pluripotency markers (Oct-4, Nanog), and differentiation markers (Brachyury, Nestin).
- Assessment of ligand domain integrity upon surface adsorption.
Main Results:
- The FIII9'10-dimer supported the highest number of mES cell colonies compared to monomer, trimer, tetramer, and gelatin.
- mES cell morphology transitioned from dispersed to tight with increasing leukemia inhibitory factor (LIF) concentrations on FIII9'10-dimer surfaces.
- In the presence of LIF, FIII9'10-dimer surfaces induced transient upregulation of Oct-4, Brachyury, and Nestin, while Nanog upregulation maintained pluripotency.
- No evidence of fibronectin domain unfolding upon surface adsorption was observed.
Conclusions:
- Dimeric integrin alpha5beta1 ligands significantly enhance mES cell adhesion and colony formation.
- mES cell behavior on dimeric ligands is primarily a morphological phenomenon influenced by LIF.
- These findings provide a foundation for controlling ES cell fate through synthetic substrates and integrin ligand presentation in tissue engineering.
