High Macromolecular Crowding in Liposomes from Microfluidics

Advanced Science, vol. 9, pp. e2201169

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.

Authors 7

1. Luis P. B. Guerzoni Aachen DWI - Leibniz Institute for Interactive Materials

   DWI – Leibniz Institute for Interactive Materials

   Affiliation as printed

   DWI‐Leibniz Institute for Interactive Materials Forckenbeckstrasse 50 52074 Aachen Germany

   DWI-Leibniz Institute for Interactive Materials, Forckenbeckstrasse 50, 52074 Aachen, Germany

2. André V. C. de Goes Aachen Institute of Technical and Macromolecular Chemistry DWI - Leibniz Institute for Interactive Materials

   DWI – Leibniz Institute for Interactive Materials · RWTH Aachen University

   Affiliation as printed

   DWI‐Leibniz Institute for Interactive Materials Forckenbeckstrasse 50 52074 Aachen Germany

   Institute of Technical and Macromolecular Chemistry RWTH Aachen University Worringerweg 1 52074 Aachen Germany

   DWI-Leibniz Institute for Interactive Materials, Forckenbeckstrasse 50, 52074 Aachen, Germany

   Institute of Technical and Macromolecular Chemistry, RWTH Aachen University, Worringerweg 1, 52074 Aachen, Germany

3. Milara Kalacheva Aachen Institute of Technical and Macromolecular Chemistry DWI - Leibniz Institute for Interactive Materials

   DWI – Leibniz Institute for Interactive Materials · RWTH Aachen University

   Affiliation as printed

   DWI‐Leibniz Institute for Interactive Materials Forckenbeckstrasse 50 52074 Aachen Germany

   Institute of Technical and Macromolecular Chemistry RWTH Aachen University Worringerweg 1 52074 Aachen Germany

   DWI-Leibniz Institute for Interactive Materials, Forckenbeckstrasse 50, 52074 Aachen, Germany

   Institute of Technical and Macromolecular Chemistry, RWTH Aachen University, Worringerweg 1, 52074 Aachen, Germany

4. Jakub Haduła Aachen Institute of Technical and Macromolecular Chemistry DWI - Leibniz Institute for Interactive Materials

   DWI – Leibniz Institute for Interactive Materials · RWTH Aachen University

   Affiliation as printed

   DWI‐Leibniz Institute for Interactive Materials Forckenbeckstrasse 50 52074 Aachen Germany

   Institute of Technical and Macromolecular Chemistry RWTH Aachen University Worringerweg 1 52074 Aachen Germany

   DWI-Leibniz Institute for Interactive Materials, Forckenbeckstrasse 50, 52074 Aachen, Germany

   Institute of Technical and Macromolecular Chemistry, RWTH Aachen University, Worringerweg 1, 52074 Aachen, Germany

5. Matthias Mork Aachen Institute of Technical and Macromolecular Chemistry DWI - Leibniz Institute for Interactive Materials

   DWI – Leibniz Institute for Interactive Materials · RWTH Aachen University

   Affiliation as printed

   DWI‐Leibniz Institute for Interactive Materials Forckenbeckstrasse 50 52074 Aachen Germany

   Institute of Technical and Macromolecular Chemistry RWTH Aachen University Worringerweg 1 52074 Aachen Germany

   DWI-Leibniz Institute for Interactive Materials, Forckenbeckstrasse 50, 52074 Aachen, Germany

   Institute of Technical and Macromolecular Chemistry, RWTH Aachen University, Worringerweg 1, 52074 Aachen, Germany

6. Laura De Laporte Aachen Department Advanced Materials for Biomedicine Institute of Applied Medical Engineering Institute of Technical and Macromolecular Chemistry DWI - Leibniz Institute for Interactive Materials Department Advanced Materials for Biomedicine Institute of Applied Medical Engineering

   DWI – Leibniz Institute for Interactive Materials · Universitätsklinikum Aachen · RWTH Aachen University

   Affiliation as printed

   DWI‐Leibniz Institute for Interactive Materials Forckenbeckstrasse 50 52074 Aachen Germany

   Department Advanced Materials for Biomedicine Institute of Applied Medical Engineering University Hospital RWTH Aachen Forckenbeckstrasse 55 52074 Aachen Germany

   Institute of Technical and Macromolecular Chemistry RWTH Aachen University Worringerweg 1 52074 Aachen Germany

   DWI-Leibniz Institute for Interactive Materials, Forckenbeckstrasse 50, 52074 Aachen, Germany

   Department Advanced Materials for Biomedicine, Institute of Applied Medical Engineering, University Hospital RWTH Aachen, Forckenbeckstrasse 55, 52074 Aachen, Germany

   Institute of Technical and Macromolecular Chemistry, RWTH Aachen University, Worringerweg 1, 52074 Aachen, Germany

7. Arnold J. Boersma corresponding Aachen DWI - Leibniz Institute for Interactive Materials

   DWI – Leibniz Institute for Interactive Materials

   Affiliation as printed

   DWI‐Leibniz Institute for Interactive Materials Forckenbeckstrasse 50 52074 Aachen Germany

   DWI-Leibniz Institute for Interactive Materials, Forckenbeckstrasse 50, 52074 Aachen, Germany