Exploiting additive and subtractive patterning for spatially controlled and robust bacterial co-cultures

Abstract

The ability to spatially pattern bacterial co-cultures brings us closer to design a cellular environment emulating the bacterial heterogeneity in natural systems. Current bacterial patterning methods, however, can only pattern single bacterial cell types. Here, additive and subtractive microcontact printing (mCP), which was used in combination with surface chemistry to control bacterial adhesion, are compared and contrasted as to their ability to produce bacterial co-cultures. Using mannosideterminated self-assembled monolayers as the adhesive surface, we demonstrate that the kinetics of bacterial adhesion is a key determinant of pattern integrity and robustness. Based on such considerations, we have established a straightforward approach based on subtractive mCP to produce spatially controlled and robust bacterial co-cultures that are viable and resist detachment due to shear, which we believe can be extensively adopted for important cell-cell communication studies such as horizontal gene transfer and quorum sensing.

An Electrically Reversible Switchable Surface to Control and Study Early Bacterial Adhesion Dynamics in Real‐Time

“…As highlighted in the literature, [29] and demonstrated by our work, [7b] the bioinertness of the SAM depends on the properties of the bacterial species, with the hydrophobicity of bacteria playing a role. The hydrophobic marine bacterium M. hydrocarbonoclasticus exhibited the lowest adhesion on the most hydrophobic surface, while readily and firmly attaching to both positively and negatively charged surfaces.…”

“…[5] Researchers have previously fabricated surfaces, typically using self-assembled monolayers (SAMs) [6] in an attempt to control and understand cell adhesion, via the introduction of specific terminal moieties to the SAM to elicit specific surface properties. [7] However, despite the advancements achieved by investigating the interface as a ‘static’ environment, the mechanism behind the cell adhesion process remain unclear [8] representing one of the biggest challenges for many scientific areas ranging from tissue engineering, [9] medicine, [10] cell biology, [11] immunology [12] and marine biofouling. [13] In particular, researchers have tended to use cells to classify surfaces ( i.e.…”

“…In our previous study, [7b] we employed several SAMs that possessed not only different backbones (i.e., hydrophilic and hydrophobic), but also different terminal functional groups (hydrophilic, hydrophobic, positively charged, negatively charged and neutral) to study in real-time the initial stages of bacterial adhesion to surfaces. As highlighted in the literature, [29] and demonstrated by our work, [7b] the bioinertness of the SAM depends on the properties of the bacterial species, with the hydrophobicity of bacteria playing a role.…”

Application of nanotechnology to control bacterial adhesion and patterning on material surfaces

“…With shear force, the lectin domain FimH becomes separated from the main protein and allows it to switch from a low-affinity to a high-affinity state. This specific shear stress-enhanced adhesion of bacteria to mannosylated surfaces is useful for bacterial adhesion studies as it can allow the micropatterned bacterial co-cultures to be exposed to shear forces resulting from fluid flow conditions without dislodging the bacteria or causing mixing of the bacterial strains [ 45 ].…”

Microstencils to generate defined, multi-species patterns of bacteria

“…11 An alternate approach to bacterial patterning is to use surface chemical modification to immobilize bacteria. This has been shown for E. coli systems 12,13 and non-model organisms, 14 but can significantly alter cell viability and development. Another approach to patterning is direct printing (ink jet), which can produce spot patterns with a resolution of approximately 200 lm.…”