StemBond hydrogels control the mechanical microenvironment for pluripotent stem cells.

Studies of mechanical signalling are typically performed by comparing cells cultured on soft and stiff hydrogel-based substrates. However, it is challenging to independently and robustly control both substrate stiffness and extracellular matrix tethering to substrates, making matrix tethering a potentially confounding variable in mechanical signalling investigations. Moreover, unstable matrix tethering can lead to poor cell attachment and weak engagement of cell adhesions. To address this, we developed StemBond hydrogels, a hydrogel in which matrix tethering is robust and can be varied independently of stiffness. We validate StemBond hydrogels by showing that they provide an optimal system for culturing mouse and human pluripotent stem cells. We further show how soft StemBond hydrogels modulate stem cell function, partly through stiffness-sensitive ERK signalling. Our findings underline how substrate mechanics impact mechanosensitive signalling pathways regulating self-renewal and differentiation, indicating that optimising the complete mechanical microenvironment will offer greater control over stem cell fate specification.

A biomechanical switch regulates the transition towards homeostasis in oesophageal epithelium.

Epithelial cells rapidly adapt their behaviour in response to increasing tissue demands. However, the processes that finely control these cell decisions remain largely unknown. The postnatal period covering the transition between early tissue expansion and the establishment of adult homeostasis provides a convenient model with which to explore this question. Here, we demonstrate that the onset of homeostasis in the epithelium of the mouse oesophagus is guided by the progressive build-up of mechanical strain at the organ level. Single-cell RNA sequencing and whole-organ stretching experiments revealed that the mechanical stress experienced by the growing oesophagus triggers the emergence of a bright Krüppel-like factor 4 (KLF4) committed basal population, which balances cell proliferation and marks the transition towards homeostasis in a yes-associated protein (YAP)-dependent manner. Our results point to a simple mechanism whereby mechanical changes experienced at the whole-tissue level are integrated with those sensed at the cellular level to control epithelial cell fate.

Equally probable positive and negative Poisson's ratios in disordered planar systems.

Auxetic materials, characterised by a negative Poisson's ratio, have properties that are different from most conventional materials. These are a result of the constraints on the kinematics of the material's basic structural components, and have important technological implications. Models of these materials have been studied extensively, but theoretical descriptions have remained largely limited to materials with an ordered microstructure. Here we investigate whether negative Poisson's ratios can arise spontaneously in disordered systems. To this end, we develop a quantitative description of the structure in systems of connected basic elements, which enables us to analyse the local and global responses to small external tensile forces. We find that the Poisson's ratios in these disordered systems are equally likely to be positive or negative on both the element and system scales. Separating the strain into translational, rotational and expansive components, we find that the translational strains of neighbouring basic structural elements are positively correlated, while their rotations are negatively correlated. There is no correlation in this type of system between the local auxeticity and local structural characteristics. Our results suggest that auxeticity is more common in disordered structures than the ubiquity of positive Poisson's ratios in macroscopic materials would suggest.

A microfluidic device for characterizing nuclear deformations.

Cell nuclei experience and respond to a wide range of forces, both in vivo and in vitro. In order to characterize the nuclear response to physical stress, we developed a microfluidic chip and used it to apply mechanical stress to live cells and measure their nuclear deformability. The device design is optimized for the detection of both nucleus and cytoplasm, which can then be conveniently quantified using a custom-written Matlab program. We measured nuclear sizes and strains of embryonic stem cells, for which we observed negative Poisson ratios in the nuclei. In addition, we were able to detect changes in the nuclear response after treatment with actin depolymerizing and chromatin decondensing agents. Finally, we showed that the device can be used for biologically relevant high-resolution confocal imaging of cells under compression. Thus, the device presented here allows for accurate physical phenotyping at high throughput and has the potential to be applied to a range of cell types.