Protein–Substrate Adhesion in Microcontact Printing Regulates Cell Behavior

Abstract

Microcontact printing (μCP) is widely used to create patterns of biomolecules essential for studies of cell mechanics, migration, and tissue engineering. However, different types of μCPs may create micropatterns with varied protein-substrate adhesion, which may change cell behaviors and pose uncertainty in result interpretation. Here, we characterize two μCP methods for coating extracellular matrix (ECM) proteins (stamp-off and covalent bond) and demonstrate for the first time the important role of protein-substrate adhesion in determining cell behavior. We found that, as compared to cells with weaker traction force (e.g., endothelial cells), cells with strong traction force (e.g., vascular smooth muscle cells) may delaminate the ECM patterns, which reduced cell viability as a result. Importantly, such ECM delamination was observed on patterns by stamp-off but not on the patterns by covalent bonds. Further comparisons of the displacement of the ECM patterns between the normal VSMCs and the force-reduced VSMCs suggested that the cell traction force plays an essential role in this ECM delamination. Together, our results indicated that μCPs with insufficient adhesion may lead to ECM delamination and cause cell death, providing new insight for micropatterning in cell-biomaterial interaction on biointerfaces.

Engineering the cellular mechanical microenvironment – from bulk mechanics to the nanoscale

“…Moreover, these techniques can be readily applied to hydrogels with different stiffness to combine both surface patterning and substrate stiffness. However, it has been reported that different stamping methods can result in differences in ligand-substrate adhesion, which in turn can affect cell behaviour, so special care must be taken to select the chemistry of the surface ligand to avoid masking the effect of the topography on cell behaviour (Hu et al, 2018).…”

Advances in Single-Cell Printing

“…However, cell viability could be affected in practice when stress is exercised on the cell during the transferring process. To obtain cell arrays with microcontact printing, it is more common to print cell adherent molecules onto the substrate through microcontact printing first and then apply cells on the substrate to form the cell array [59][60][61][62][63].…”

“…In contact printing of cells, cell adherent molecules such as extracellular matrix (ECM) proteins including fibronectin, laminin, and vitronectin are generally used [63,64]. These functional anchoring units are usually immobilized on surface-active molecules, such as self-assemble monolayers (SAMs) or other functional groups, which were previously printed on substrates with microcontact printing [65].…”

Continuous monitoring of cell transfection efficiency with micropatterned substrates

“…These cell arrays have been proven to be suitable for gene transfection, having the potential to overcome the limitations of conventional cell cultures (McConnell et al, 2011). Thanks to the versatility of the technique, great control over cell interactions can be achieved, including cell-material and cell-cell interactions, while also enabling the easy generation of a high number of data points in a small substrate area (Garcia-Hernando et al, 2020;Gonzalez-Pujana et al, 2019;Hamon et al, 2016;Hu et al, 2018;Kobel & Lutolf, 2010). Cell patterns can be directly transfected and monitored, without the need of detaching and transporting the cells to a different setup, as the pattern itself simplifies quantification due to the highly controlled localization of the cells.…”