Controlled packing and single-droplet resolution of 3D-printed functional synthetic tissues

Alcinesio, Alessandro; Meacock, O. J.; Allan, Rebecca G.; Mónico, Carina; Schild, Vanessa Restrepo; Cazimoglu, Idil; Cornall, Matthew T.; Kumar, Ravinash Krishna

doi:10.1038/s41467-020-15953-y

Cited by 79 publications

(112 citation statements)

References 43 publications

(51 reference statements)

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“…Undecane and silicone oil AR20 (Sigma-Aldrich) were filtered before use through 0.22-µm filters (Corning ® ) under vacuum. An optimised lipid/oil solution was prepared for printing 65 . Lipid films were prepared by bringing ampoules to room temperature and dissolving in anhydrous chloroform (Sigma-Aldrich) to make a chloroform/lipid solution at lipid concentrations of 25 mg mL −1 .…”

Section: Methodsmentioning

confidence: 99%

Droplet printing reveals the importance of micron-scale structure for bacterial ecology

et al. 2021

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Bacteria often live in diverse communities where the spatial arrangement of strains and species is considered critical for their ecology. However, a test of this hypothesis requires manipulation at the fine scales at which spatial structure naturally occurs. Here we develop a droplet-based printing method to arrange bacterial genotypes across a sub-millimetre array. We print strains of the gut bacterium Escherichia coli that naturally compete with one another using protein toxins. Our experiments reveal that toxin-producing strains largely eliminate susceptible non-producers when genotypes are well-mixed. However, printing strains side-by-side creates an ecological refuge where susceptible strains can persist in large numbers. Moving to competitions between toxin producers reveals that spatial structure can make the difference between one strain winning and mutual destruction. Finally, we print different potential barriers between competing strains to understand how ecological refuges form, which shows that cells closest to a toxin producer mop up the toxin and protect their clonemates. Our work provides a method to generate customised bacterial communities with defined spatial distributions, and reveals that micron-scale changes in these distributions can drive major shifts in ecology.

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Section: Methodsmentioning

confidence: 99%

Droplet printing reveals the importance of micron-scale structure for bacterial ecology

et al. 2021

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“…Undecane and silicone oil AR20 (Sigma-Aldrich) were filtered before use through 0.22 μm filters (Corning ® ) under vacuum. An optimised lipid/oil solution was prepared for printing 57 . Lipid films were prepared by bringing ampoules to room temperature and dissolving in anhydrous chloroform (Sigma-Aldrich) to make a chloroform/lipid solution at lipid concentrations of 25 mg mL -1 .…”

Section: Methodsmentioning

confidence: 99%

Droplet printing reveals the importance of micron-scale structure for bacterial ecology

Kumar

Meiller-Legrand

Alcinesio

et al. 2020

Preprint

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Bacteria often live in diverse communities where the spatial arrangement of strains and species is considered critical for their ecology, including whether strains can coexist, which are ecologically dominant, and how productive they are as a community. However, a test of the importance of spatial structure requires manipulation at the fine scales at which this structure naturally occurs. Here we develop a droplet-based printing method to arrange different bacterial genotypes across a sub-millimetre array. We use this to test the importance of fine-scale spatial structure by printing strains of the gut bacterium Escherichia coli that naturally compete with one another using protein toxins. This reveals that the spatial arrangement of bacterial genotypes is important for ecological outcomes. Toxin-producing strains largely eliminate susceptible non-producers when genotypes are well-mixed. However, printing strains side-by-side creates an ecological refuge such that susceptible strains can coexist with toxin producers, even to the extent that a susceptible strain outnumbers the toxin producer. Head-to-head competitions between toxin producers also reveals strong effects, where spatial structure can make the difference between one strain winning and mutual destruction. Finally, we print different potential barriers between two competing strains to understand why space is so important. This reveals the importance of processes that limit the free diffusion of molecules. Specifically, we show that cells closest to a toxin producer bind to and capture toxin molecules, which creates a refuge for their clonemates. Our work provides a new method to generate customised bacterial communities with defined spatial distributions, and reveals that micron-scale changes in these distributions can drive major shifts in their ecology.

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“…Hence, when two such compartments are brought together a lipid bilayer forms between them, termed droplet interface bilayer (DIB) 13 . Networks of droplets interconnected by DIBs can be generated manually 14,15 or by 3D-printing 12,16 . Within oil, these structures can process external light 15,17 , mechanical 18 or electrical 11 stimuli, but they cannot handle input from water-soluble signalling molecules.…”

Section: Introductionmentioning

confidence: 99%

A lipid-based parallel processor for chemical signals

Cazimoglu

Bayley

2021

Preprint

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A key goal of bottom-up synthetic biology is to construct cell- and tissue-like structures. Underpinning cellular life is the ability to process several external chemical signals, often in parallel. Until now, however, cell- and tissue-like structures have only been constructed with one signalling pathway. Here, we construct a dual-signal processor from the bottom up in a modular fashion. The processor comprises three aqueous compartments bounded by lipid bilayers and operates in an aqueous environment. It can receive two chemical signals from the external environment, process them orthogonally, and then produce a distinct output for each signal. It is suitable for both sensing and enzymatic processing of environmental signals with fluorescence and molecular outputs. In the future, such processors could serve as smart drug delivery vehicles or as modules within synthetic tissues to control their behaviour in response to external chemical signals.

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Controlled packing and single-droplet resolution of 3D-printed functional synthetic tissues

Cited by 79 publications

References 43 publications

Droplet printing reveals the importance of micron-scale structure for bacterial ecology

Droplet printing reveals the importance of micron-scale structure for bacterial ecology

Droplet printing reveals the importance of micron-scale structure for bacterial ecology

A lipid-based parallel processor for chemical signals

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