Stem Cells
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taminating undifferentiated ES cells could result in tumourigenesis.
3. Microenvironment/niche
Understanding the microenvironments in which stem cells reside and differentiate in vivo and try- ing to recapitulate these in vitro to further control stem cell differentiation has become an increasing- ly important area of stem cell research. It is clear that factors other than those added to the cell cul- ture medium can have a profound effect on stem cell self-renewal and differentiation28. Stem cell niches are 3D environments that subject cells to many different interactions which all combine to affect their fate. These include immobilised growth factors, extracellular matrix (ECM) components and neighbouring cell types, in addition to soluble growth factors. In particular, focus has been on the biochemical and mechanical influence of different ECM components and how these and the 3D con- figuration of cells affects their fate.
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The ECM signals to cells principally via integrin receptors causing changes in cell shape, migration and protein expression28. In addition, the ECM provides a physical framework, affecting the mechanical forces cells are subjected to. There are several examples of how biochemical and mechan- ical forces affect stem cell differentiation and increasingly innovative microfabrication techniques have been used to investigate these influences, allowing a high throughput and cost-effective way of discovering how different materials affect stem cell fate29. For example, different ECM and cell adhesion factors can be robotically spotted on to microarrays in various combinations, allowing screens of tens to hundreds of putative microenvi- ronments. La Flaim et al used this technique to probe interactions of ECM components in combi- nation with soluble growth factors30. A multiwell microarray platform that allows 1,200 simultane- ous experiments on 240 unique signalling environ- ments was developed. A reporter ES cell line (GFP under the control of the MHC promoter) was used to monitor cardiac differentiation using a confocal microarray scanner. The results were consistent with what has previously been published, providing proof of concept for this approach.
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The effect of mechanical forces on stem cell differentiation has also become a major topic of investigation. It is clear that applied mechanical forces can affect the activity and expression of transcription factors and chromatin remodelling enzymes in turn affecting stem cell fate. A study investigating different polyacrylamide gels showed that gel stiffness had a dramatic effect on
Drug Discovery World Winter 2011/12
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