Microfluidic production and characterization of biofunctionalized giant unilamellar vesicles for targeted intracellular cargo delivery

Oskar Staufer*, Silvia Antona, Dennis Zhang, Júlia Csatári, Martin Schröter, Jan-Willi Janiesch, Sebastian Fabritz, Imre Berger, Ilia Platzman*, Joachim P Spatz*

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)peer-review

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Abstract

Lipid-based vesicles have found widespread applications in the life sciences, allowing for fundamental insights into membrane-based processes in cell biology and as carrier systems for drug delivery purposes. So far, mostly small unilamellar vesicles (SUVs) with diameters of ~100 nm have been applied as carrier systems for biomedical applications. Despite this progress, several systematic limitations have arisen due to SUV dimensions, e.g., the size and total amount of applicable cargo is limited. Giant unilamellar vesicles (GUVs) might offer a pragmatic al-ternative for efficient cargo delivery. However, due to the lack of reliable high-throughput production technologies for GUV-carrier systems, only little is known about their interaction with cells. Here we present a microfluidic- based mechanical droplet-splitting pipeline for the production of carrier-GUVs with diameters of ~2 μm. The technology developed allows for highly efficient cargo loading and unprecedented control over the biological and physicochemical properties of GUV membranes. By generating differently charged (between −31 and + 28 mV), bioligand-conjugated (e.g. with E-cadherin, NrCam and antibodies) and PEG-conjugated GUVs, we performed a detailed investigation of attractive and repulsive GUV-cell interactions. Fine-tuning of these interactions allowed for targeted cellular GUV delivery. Moreover, we evaluated strategies for intracellular GUV cargo release by ly-sosomal escape mediated by the pH sensitive lipid DOBAQ, enabling cytoplasmic transmission. The presented GUV delivery technology and the systematic characterization of associated GUV-cell interactions could provide a means for more efficient drug administration and will pave the way for hitherto impossible approaches towards a targeted delivery of advanced cargo such as microparticles, viruses or macromolecular DNA-robots.
Original languageEnglish
Article number120203
Number of pages14
JournalBiomaterials
Volume264
Early online date9 Sep 2020
DOIs
Publication statusPublished - 1 Jan 2021

Bibliographical note

Funding Information:
The authors acknowledge funding from the Federal Ministry of Education and Research of Germany, Grant Agreement no. 13XP5073A, PolyAntiBak and the MaxSynBio Consortium, the later is jointly funded by the Federal Ministry of Education and Research of Germany and the Max Planck Society . They also acknowledge the support from the Volkswagen Stiftung (priority call ‘Life?’) and the BBSRC / EPSRC Research Centre for synthetic biology at the University of Bristol BrisSynBio (BB/L01386X/1). The authors are grateful to Andrea Hellwig (Interdisciplinary Centre for Neurosciences Heidelberg) for technical assistance during TEM preparation and imaging as well as Martin Pelosse and Julia Ricken for production and purification of baculoviruses and recombinant GFP, respectively. The authors thank the laboratory of Hilmar Bading, Department of Neurobiology and IZN, Heidelberg University, for providing primary hippocampal neurons. The authors further thank the MS Core Facility (MPI for Medical Research) for technical assistance and performance of mass spectrometry analysis. O.S. acknowledges support from the Heidelberg Biosciences International Graduate School and the Max Planck School Matter to Life . O.S. is the Meurer Visiting Professor at the Max Planck Center for Minimal Biology, Bristol. I.B. is investigator of the European Research Council (ERC-2018-ADG 834631 DNA- DOCK ). J.P.S. is the Weston Visiting Professor at the Weizmann Institute of Science and part of the excellence cluster CellNetworks at the University of Heidelberg. The Max Planck Society is appreciated for its general support.

Funding Information:
The authors acknowledge funding from the Federal Ministry of Education and Research of Germany, Grant Agreement no. 13XP5073A, PolyAntiBak and the MaxSynBio Consortium, the later is jointly funded by the Federal Ministry of Education and Research of Germany and the Max Planck Society. They also acknowledge the support from the Volkswagen Stiftung (priority call ?Life??) and the BBSRC/EPSRC Research Centre for synthetic biology at the University of Bristol BrisSynBio (BB/L01386X/1). The authors are grateful to Andrea Hellwig (Interdisciplinary Centre for Neurosciences Heidelberg) for technical assistance during TEM preparation and imaging as well as Martin Pelosse and Julia Ricken for production and purification of baculoviruses and recombinant GFP, respectively. The authors thank the laboratory of Hilmar Bading, Department of Neurobiology and IZN, Heidelberg University, for providing primary hippocampal neurons. The authors further thank the MS Core Facility (MPI for Medical Research) for technical assistance and performance of mass spectrometry analysis. O.S. acknowledges support from the Heidelberg Biosciences International Graduate School and the Max Planck School Matter to Life. O.S. is the Meurer Visiting Professor at the Max Planck Center for Minimal Biology, Bristol. I.B. is investigator of the European Research Council (ERC-2018-ADG 834631 DNA-DOCK). J.P.S. is the Weston Visiting Professor at the Weizmann Institute of Science and part of the excellence cluster CellNetworks at the University of Heidelberg. The Max Planck Society is appreciated for its general support.

Publisher Copyright:
© 2020 The Authors

Keywords

  • Giant unilamellar vesicles
  • Targeted delivery
  • GUV-cell interactions
  • Microfluidics

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