High Macromolecular Crowding in Liposomes from Microfluidics

Luis P. B. Guerzoni; André V. C. de Goes; Milara Kalacheva; Jakub Haduła; Matthias Mork; Laura De Laporte; Arnold J. Boersma

doi:10.1002/advs.202201169

High Macromolecular Crowding in Liposomes from Microfluidics

Luis P. B. Guerzoni, André V. C. de Goes, Milara Kalacheva, Jakub Haduła, Matthias Mork, Laura De Laporte, Arnold J. Boersma

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Research output: Contribution to journal › Article › Academic › peer-review

Abstract

The intracellular environment is crowded with macromolecules that influence biochemical equilibria and biomacromolecule diffusion. The incorporation of such crowding in synthetic cells would be needed to mimic the biochemistry of living cells. However, only a few methods provide crowded artificial cells, moreover providing cells with either heterogeneous size and composition or containing a significant oil fraction. Therefore, a method that generates monodisperse liposomes with minimal oil content and tunable macromolecular crowding using polydimethylsiloxane (PDMS)-based microfluidics is presented. Lipid stabilized water-in-oil-in-water emulsions that are stable for at least several months and with a high macromolecular crowder concentration that can be controlled with the external osmolality are formed. A crucial feature is that the oil phase can be removed using high flow conditions at any point after production, providing the highly crowded liposomes. Genetically encoded macromolecular crowding sensors show that the high level of macromolecular crowding in the emulsions is fully retained throughout the generation of minimal-oil lipid bilayers. This modular and robust platform will serve the study of biochemistry under physiologically relevant crowding conditions.

Original language	English
Article number	2201169
Pages (from-to)	1-12
Journal	Advanced Science
Volume	9
Issue number	27
DOIs	https://doi.org/10.1002/advs.202201169
Publication status	Published - 23 Sept 2022
Externally published	Yes

Bibliographical note

Funding Information:
L.P.B.G. and A.V.C.G. contributed equally to this work. The authors thank Mr. Fabian Schmitz for preparing schematic representations. This work was supported by The European Union (European Research Council Consolidator Grant PArtCell No. 864528). Parts of the analytical investigations were performed at the Center for Chemical Polymer Technology CPT, which was supported by the European Commission and the federal state of North Rhine-Westphalia (No. 300088302).

Funding Information:
L.P.B.G. and A.V.C.G. contributed equally to this work. The authors thank Mr. Fabian Schmitz for preparing schematic representations. This work was supported by The European Union (European Research Council Consolidator Grant PArtCell No. 864528). Parts of the analytical investigations were performed at the Center for Chemical Polymer Technology CPT, which was supported by the European Commission and the federal state of North Rhine‐Westphalia (No. 300088302).

Publisher Copyright:
© 2022 The Authors. Advanced Science published by Wiley-VCH GmbH.

Keywords

FRET sensors
macromolecular crowding
microfluidics
sartificial cells

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Advanced Science - 2022 - Guerzoni - High Macromolecular Crowding in Liposomes from MicrofluidicsFinal published version, 4.37 MBLicence: CC BY

Cite this

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title = "High Macromolecular Crowding in Liposomes from Microfluidics",

abstract = "The intracellular environment is crowded with macromolecules that influence biochemical equilibria and biomacromolecule diffusion. The incorporation of such crowding in synthetic cells would be needed to mimic the biochemistry of living cells. However, only a few methods provide crowded artificial cells, moreover providing cells with either heterogeneous size and composition or containing a significant oil fraction. Therefore, a method that generates monodisperse liposomes with minimal oil content and tunable macromolecular crowding using polydimethylsiloxane (PDMS)-based microfluidics is presented. Lipid stabilized water-in-oil-in-water emulsions that are stable for at least several months and with a high macromolecular crowder concentration that can be controlled with the external osmolality are formed. A crucial feature is that the oil phase can be removed using high flow conditions at any point after production, providing the highly crowded liposomes. Genetically encoded macromolecular crowding sensors show that the high level of macromolecular crowding in the emulsions is fully retained throughout the generation of minimal-oil lipid bilayers. This modular and robust platform will serve the study of biochemistry under physiologically relevant crowding conditions.",

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note = "Funding Information: L.P.B.G. and A.V.C.G. contributed equally to this work. The authors thank Mr. Fabian Schmitz for preparing schematic representations. This work was supported by The European Union (European Research Council Consolidator Grant PArtCell No. 864528). Parts of the analytical investigations were performed at the Center for Chemical Polymer Technology CPT, which was supported by the European Commission and the federal state of North Rhine-Westphalia (No. 300088302). Funding Information: L.P.B.G. and A.V.C.G. contributed equally to this work. The authors thank Mr. Fabian Schmitz for preparing schematic representations. This work was supported by The European Union (European Research Council Consolidator Grant PArtCell No. 864528). Parts of the analytical investigations were performed at the Center for Chemical Polymer Technology CPT, which was supported by the European Commission and the federal state of North Rhine‐Westphalia (No. 300088302). Publisher Copyright: {\textcopyright} 2022 The Authors. Advanced Science published by Wiley-VCH GmbH.",

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AU - Boersma, Arnold J.

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N2 - The intracellular environment is crowded with macromolecules that influence biochemical equilibria and biomacromolecule diffusion. The incorporation of such crowding in synthetic cells would be needed to mimic the biochemistry of living cells. However, only a few methods provide crowded artificial cells, moreover providing cells with either heterogeneous size and composition or containing a significant oil fraction. Therefore, a method that generates monodisperse liposomes with minimal oil content and tunable macromolecular crowding using polydimethylsiloxane (PDMS)-based microfluidics is presented. Lipid stabilized water-in-oil-in-water emulsions that are stable for at least several months and with a high macromolecular crowder concentration that can be controlled with the external osmolality are formed. A crucial feature is that the oil phase can be removed using high flow conditions at any point after production, providing the highly crowded liposomes. Genetically encoded macromolecular crowding sensors show that the high level of macromolecular crowding in the emulsions is fully retained throughout the generation of minimal-oil lipid bilayers. This modular and robust platform will serve the study of biochemistry under physiologically relevant crowding conditions.

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High Macromolecular Crowding in Liposomes from Microfluidics

Abstract

Bibliographical note

Keywords

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