Person: Baker, Brendon M.
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Baker
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Brendon M.
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Baker, Brendon M.
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Publication Micropatterned Multicolor Dynamically Adhesive Substrates to Control Cell Adhesion and Multicellular Organization(American Chemical Society, 2014) Rodriguez, Natalia M.; Desai, Ravi A.; Trappmann, Britta; Baker, Brendon M.; Chen, ChristopherWe present a novel technique to examine cell–cell interactions and directed cell migration using micropatterned substrates of three distinct regions: an adhesive region, a nonadhesive region, and a dynamically adhesive region switched by addition of a soluble factor to the medium. Combining microcontact printing with avidin–biotin capture chemistry, we pattern nonadhesive regions of avidin that become adhesive through the capture of biotinylated fibronectin. Our strategy overcomes several limitations of current two-color dynamically adhesive substrates by incorporating a third, permanently nonadhesive region. Having three spatially and functionally distinct regions allows for the realization of more complex configurations of cellular cocultures as well as intricate interface geometries between two cell populations for diverse heterotypic cell–cell interaction studies. We can now achieve spatial control over the path and direction of migration in addition to temporal control of the onset of migration, enabling studies that better recapitulate coordinated multicellular migration and organization in vitro. We confirm that cellular behavior is unaltered on captured biotinylated fibronectin as compared to printed fibronectin by examining the cells’ ability to spread, form adhesions, and migrate. We demonstrate the versatility of this approach in studies of migration and cellular cocultures, and further highlight its utility by probing Notch–Delta juxtacrine signaling at a patterned interface.Publication A DNA-based molecular probe for optically reporting cellular traction forces(2014) Blakely, Brandon L.; Dumelin, Christoph E.; Trappmann, Britta; McGregor, Lynn M.; Choi, Colin K.; Anthony, Peter C.; Duesterberg, Van K.; Baker, Brendon M.; Block, Steven M.; Liu, David; Chen, Christopher S.We developed molecular tension probes (TPs) that report traction forces of adherent cells with high spatial resolution, can be linked to virtually any surface, and obviate monitoring deformations of elastic substrates. TPs consist of DNA hairpins conjugated to fluorophore-quencher pairs that unfold and fluoresce when subjected to specific forces. We applied TPs to reveal that cellular traction forces are heterogeneous within focal adhesions and localized at their distal edges.Publication Differentiation alters stem cell nuclear architecture, mechanics, and mechano-sensitivity(eLife Sciences Publications, Ltd, 2016) Heo, Su-Jin; Driscoll, Tristan P; Thorpe, Stephen D; Nerurkar, Nandan L; Baker, Brendon M.; Yang, Michael T; Chen, Christopher; Lee, David A; Mauck, Robert LMesenchymal stem cell (MSC) differentiation is mediated by soluble and physical cues. In this study, we investigated differentiation-induced transformations in MSC cellular and nuclear biophysical properties and queried their role in mechanosensation. Our data show that nuclei in differentiated bovine and human MSCs stiffen and become resistant to deformation. This attenuated nuclear deformation was governed by restructuring of Lamin A/C and increased heterochromatin content. This change in nuclear stiffness sensitized MSCs to mechanical-loading-induced calcium signaling and differentiated marker expression. This sensitization was reversed when the ‘stiff’ differentiated nucleus was softened and was enhanced when the ‘soft’ undifferentiated nucleus was stiffened through pharmacologic treatment. Interestingly, dynamic loading of undifferentiated MSCs, in the absence of soluble differentiation factors, stiffened and condensed the nucleus, and increased mechanosensitivity more rapidly than soluble factors. These data suggest that the nucleus acts as a mechanostat to modulate cellular mechanosensation during differentiation. DOI: http://dx.doi.org/10.7554/eLife.18207.001