Assembly of biomimetic microreactors using caged-coacervate droplets

Jobdeedamrong, Arjaree; Cao, Shoupeng; Harley, Iain; Crespy, Daniel; Landfester, Katharina; Silva, Lucas Caire da

doi:10.1039/d2nr05101j

Cited by 12 publications

(11 citation statements)

References 39 publications

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“…Different strategies have been explored to stabilize complex coacervates, including electrostatic repulsion -by altering the positive to negative molar charge ratio (and hence the excess surface charge on the droplets) -or, more intriguingly, dispersion of the bulk coacervate phase (collected after centrifugation) into Oligopeptides PAH/pGlu [148] Polyelectrolytes PDDA/PAA [254] Protein-polymer conjugates Q-Am/Su-Am [253] Block copolymers Q-Am/CM-Am [129,[255][256][257][258] Small unilamellar vesicles Spermine/polyU [165] PDDA/PAA, PAH/PAA, PAH/ADP [235] PDDA/ATP, PDDA/PAA, PEI/ATP [236] Nanoparticles PDDA/ATP, PDDA/PAA [237] PDDA/CM-dex [238] Q-Am/CM-Am [239] Microgels Polyampholyte [240] Cell fragments Q-Am/HA [241] DEAE-dex/DNA [242] Bacteria PDDA/ATP [243] Organelle confinement Proteinosome Fatty acids [150] Polymersome Q-Am/CM-dex [256] Colloidosome PDDA/CM-dex [238] Chloroplast Q-Am/HA [241] PDDA/CM-dex [265] Bacteria Q-Am/HA [241] PDDA/ATP [243] Cytoskeleton self-assembly FtsZ pLys/RNA [202] pLys/GTP [203] Actin pLys/pGlu [137] FUS [200] DNA nanotube PDDA/PAA [149] a ) See legend of Table 1, PEI: polyethyleneimine, HA: sodium hyaluronate, PEAD: poly(ethylene argininylaspartate diglyceride), Su-Am: succinyl amylose, pGlu: poly-l-glutamic acid, GTP: guanosine triphosphate.…”

Section: Of the Thermodynamic Instability Of Coacervate Dropletsmentioning

confidence: 99%

“…nano-or microscale objects (Figure 8a), such as liposomes (small unilamellar vesicles), [165,235,236] nanoparticles, [237][238][239] microgels, [240] cell fragments, [241,242] or even living cells; [243] ii) amphiphilic (macro)molecules (Figure 8b), including fatty acids, [244,245] phospholipids, [246][247][248][249][250] surfactants, [251,252] proteinpolymer conjugates, [253] self-assembling oligopeptides, [148] comb polyelectrolytes, [254] or block copolymers. [255][256][257][258] Depending on the nature of the membrane-forming components, different mechanisms may explain their interfacial adsorption on coacervates.…”

Section: Coacervate-guided Membrane Self-assemblymentioning

confidence: 99%

“…Nano-or microscale stabilizers typically result in shells with relatively large pores, allowing the unrestricted diffusion of both small and large molecules. [165,239] Surprisingly, lipid-and polymer-coated coacervates have been shown to retain their capacity to spontaneously accumulate small molecules and ions, such as glucose, hydrogen peroxide, or magnesium ions, but also larger macromolecules, including dextran chains, proteins, RNA, and DNA, suggesting the existence of defects/pores in the membrane. [246,247,257] These defects/pores can be viewed as either detrimental (as they allow unrestricted diffusion of both small and large solutes) or advantageous (since they facilitate solute exchange with the environment) depending on the considered function.…”

Section: Coacervate-guided Membrane Self-assemblymentioning

confidence: 99%

“…A broad palette of building blocks has been reported to induce membranization of coacervate droplets. Schematically, these can be divided into two main classes, namely, i) nano‐ or microscale objects (Figure 8a), such as liposomes (small unilamellar vesicles), [ 165,235,236 ] nanoparticles, [ 237–239 ] microgels, [ 240 ] cell fragments, [ 241,242 ] or even living cells; [ 243 ] ii) amphiphilic (macro)molecules (Figure 8b), including fatty acids, [ 244,245 ] phospholipids, [ 246–250 ] surfactants, [ 251,252 ] protein–polymer conjugates, [ 253 ] self‐assembling oligopeptides, [ 148 ] comb polyelectrolytes, [ 254 ] or block copolymers. [ 255–258 ]…”

Section: Coacervates As Cytosol‐like Templates For Synthetic Cellsmentioning

confidence: 99%

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Coacervate Droplets for Synthetic Cells

2023

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The design and construction of synthetic cells – human‐made microcompartments that mimic features of living cells – have experienced a real boom in the past decade. While many efforts have been geared toward assembling membrane‐bounded compartments, coacervate droplets produced by liquid–liquid phase separation have emerged as an alternative membrane‐free compartmentalization paradigm. Here, the dual role of coacervate droplets in synthetic cell research is discussed: encapsulated within membrane‐enclosed compartments, coacervates act as surrogates of membraneless organelles ubiquitously found in living cells; alternatively, they can be viewed as crowded cytosol‐like chassis for constructing integrated synthetic cells. After introducing key concepts of coacervation and illustrating the chemical diversity of coacervate systems, their physicochemical properties and resulting bioinspired functions are emphasized. Moving from suspensions of free floating coacervates, the two nascent roles of these droplets in synthetic cell research are highlighted: organelle‐like modules and cytosol‐like templates. Building the discussion on recent studies from the literature, the potential of coacervate droplets to assemble integrated synthetic cells capable of multiple life‐inspired functions is showcased. Future challenges that are still to be tackled in the field are finally discussed.

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Section: Of the Thermodynamic Instability Of Coacervate Dropletsmentioning

confidence: 99%

Section: Coacervate-guided Membrane Self-assemblymentioning

confidence: 99%

Section: Coacervate-guided Membrane Self-assemblymentioning

confidence: 99%

Section: Coacervates As Cytosol‐like Templates For Synthetic Cellsmentioning

confidence: 99%

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Coacervate Droplets for Synthetic Cells

2023

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“…In the past few decades, droplet microfluidics technology has developed rapidly, and it has been used in the design of microreactors because of its strong controllability in performing various operations ( Chen et al, 2022 ; Jobdeedamrong et al, 2023 ). Due to their compatibility with many chemical and biological reagents, droplet microfluidic systems have been used to prepare complex reactants and special materials, such as polymer particles ( Juang and Chiu, 2022 ), microgel particles ( Rojek et al, 2022 ), and nanoparticles ( Küçüktürkmen et al, 2022 ; Maged et al, 2022 ; Niculescu et al, 2022 ).…”

Section: Droplet Generation and Single-cell Encapsulationmentioning

confidence: 99%

Droplets microfluidics platform—A tool for single cell research

Cheng

et al. 2023

Front. Bioeng. Biotechnol.

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Cells are the most basic structural and functional units of living organisms. Studies of cell growth, differentiation, apoptosis, and cell-cell interactions can help scientists understand the mysteries of living systems. However, there is considerable heterogeneity among cells. Great differences between individuals can be found even within the same cell cluster. Cell heterogeneity can only be clearly expressed and distinguished at the level of single cells. The development of droplet microfluidics technology opens up a new chapter for single-cell analysis. Microfluidic chips can produce many nanoscale monodisperse droplets, which can be used as small isolated micro-laboratories for various high-throughput, precise single-cell analyses. Moreover, gel droplets with good biocompatibility can be used in single-cell cultures and coupled with biomolecules for various downstream analyses of cellular metabolites. The droplets are also maneuverable; through physical and chemical forces, droplets can be divided, fused, and sorted to realize single-cell screening and other related studies. This review describes the channel design, droplet generation, and control technology of droplet microfluidics and gives a detailed overview of the application of droplet microfluidics in single-cell culture, single-cell screening, single-cell detection, and other aspects. Moreover, we provide a recent review of the application of droplet microfluidics in tumor single-cell immunoassays, describe in detail the advantages of microfluidics in tumor research, and predict the development of droplet microfluidics at the single-cell level.

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Continuous Transformation from Membrane‐Less Coacervates to Membranized Coacervates and Giant Vesicles: Toward Multicompartmental Protocells with Complex (Membrane) Architectures

Zhou,

Zhang,

Moreno

et al. 2024

Angewandte Chemie

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The membranization of membrane‐less coacervates paves the way for the exploitation of complex protocells with regard to structural and cell‐like functional behaviors. However, the controlled transformation from membranized coacervates to vesicles remains a challenge. This can provide stable (multi)phase and (multi)compartmental architectures through the reconfiguration of coacervate droplets in the presence of (bioactive) polymers, bio(macro)molecules and/or nanoobjects. Herein, we present a continuous protocell transformation from membrane‐less coacervates to membranized coacervates and, ultimately, to giant hybrid vesicles. This transformation process is orchestrated by altering the balance of non‐covalent interactions through varying concentrations of an anionic terpolymer, leading to dynamic processes such as spontaneous membranization of terpolymer nanoparticles at the coacervate surface, disassembly of the coacervate phase mediated by the excess anionic charge, and the redistribution of coacervate components in membrane. The diverse protocells during the transformation course provide distinct structural features and molecular permeability. Notably, the introduction of multiphase coacervates in this continuous transformation process signifies advancements toward the creation of synthetic cells with different diffusible compartments. Our findings emphasize the highly controlled continuous structural reorganization of coacervate protocells and represents a novel step toward the development of advanced and sophisticated synthetic protocells with more precise compositions and complex (membrane) structures.

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Assembly of biomimetic microreactors using caged-coacervate droplets

Abstract: Complex coacervates are liquid-like droplets that can be used to create adaptive cell-like compartments. These compartments offer a versatile platform for the construction of bioreactors inspired by living cells. However,...

Cited by 12 publications

References 39 publications

Coacervate Droplets for Synthetic Cells

Coacervate Droplets for Synthetic Cells

Droplets microfluidics platform—A tool for single cell research

Continuous Transformation from Membrane‐Less Coacervates to Membranized Coacervates and Giant Vesicles: Toward Multicompartmental Protocells with Complex (Membrane) Architectures

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