The Journal of Membrane Biology

, Volume 248, Issue 5, pp 827–835 | Cite as

Optimization of the Electroformation of Giant Unilamellar Vesicles (GUVs) with Unsaturated Phospholipids

Article
First Online:
Received:
Accepted:

Abstract

Giant unilamellar vesicles (GUV) are widely used cell membrane models. GUVs have a cell-like diameter and contain the same phospholipids that constitute cell membranes. The most frequently used protocol to obtain these vesicles is termed electroformation, since key steps of this protocol consist in the application of an electric field to a phospholipid deposit. The potential oxidation of unsaturated phospholipids due to the application of an electric field has not yet been considered even though the presence of oxidized lipids in the membrane of GUVs could impact their permeability and their mechanical properties. Thanks to mass spectrometry analyses, we demonstrated that the electroformation technique can cause the oxidation of polyunsaturated phospholipids constituting the vesicles. Then, using flow cytometry, we showed that the amplitude and the duration of the electric field impact the number and the size of the vesicles. According to our results, the oxidation level of the phospholipids increases with their level of unsaturation as well as with the amplitude and the duration of the electric field. However, when the level of lipid oxidation exceeds 25 %, the diameter of the vesicles is decreased and when the level of lipid oxidation reaches 40 %, the vesicles burst or reorganize and their rate of production is reduced. In conclusion, the classical electroformation method should always be optimized, as a function of the phospholipid used, especially for producing giant liposomes of polyunsaturated phospholipids to be used as a cell membrane model.

Keywords

Electroformation Giant unilamellar vesicles Phospholipid Oxidation Electric field Membrane model 
This is a preview of subscription content, log in to check access.

Notes

Acknowledgments

The authors thank the « Agence Nationale de la Recherche » programs Oxylipi2 (ANR-13-ASTR-0029), and Memove (ANR-11-BS01-006) and the « Agence Nationale de Sécurité Sanitaire de l’Alimentation, de l’Environnement et du Travail ANSES » program MARFEM (EST-12-188) for funding. This research was conducted in the scope of the European associated laboratories EBAM and is partly funded by the CNRS, the University of Paris-Sud, Gustave Roussy, and the EDF Foundation.

Supplementary material

232_2015_9828_MOESM1_ESM.doc (106 kb)
Supplementary material 1 (DOC 106 kb)

References

  1. Angelova MI (2007) Perspectives in supramolecular chemistry, vol 6., Giant VesiclesWiley, Chichester, pp 26–36CrossRefGoogle Scholar
  2. Angelova MI, Dimitrov DS (1986) Liposome electroformation. Faraday Discuss 81:303. doi: 10.1039/dc9868100303 CrossRefGoogle Scholar
  3. Angelova MI, Dimitrov DS (1988) Trends in colloid and interface science II, vol 76. Springer, Berlin, pp 59–67. doi: 10.1007/BFb0114157 CrossRefGoogle Scholar
  4. Bagatolli LA, Needham D (2014) Quantitative optical microscopy and micromanipulation studies on the lipid bilayer membranes of giant unilamellar vesicles. Chem Phys Lipids 181:99–120. doi: 10.1016/j.chemphyslip.2014.02.009 CrossRefPubMedGoogle Scholar
  5. Breton M, Delemotte L, Silve A et al (2012) Transport of siRNA through lipid membranes driven by nanosecond electric pulses: an experimental and computational study. J Am Chem Soc 134:13938–13941. doi: 10.1021/ja3052365 CrossRefPubMedGoogle Scholar
  6. Dimova R (2014) Recent developments in the field of bending rigidity measurements on membranes. Adv Colloid Interface Sci 208:225–234. doi: 10.1016/j.cis.2014.03.003 CrossRefPubMedGoogle Scholar
  7. Dimova R, Aranda S, Bezlyepkina N et al (2006) A practical guide to giant vesicles. Probing the membrane nanoregime via optical microscopy. J Phys Condens Matter 18:S1151–S1176. doi: 10.1088/0953-8984/18/28/S04 CrossRefPubMedGoogle Scholar
  8. Fenz SF, Sengupta K (2012) Giant vesicles as cell models. Integr Biol (Camb) 4:982–995. doi: 10.1039/c2ib00188h CrossRefGoogle Scholar
  9. Halliwell B, Chirico S (1993) Lipid peroxidation: its mechanism, measurement, and significance. Am J Clin Nutr 57:715S–724SPubMedGoogle Scholar
  10. Mauroy C, Portet T, Winterhalder M et al (2012) Giant lipid vesicles under electric field pulses assessed by non invasive imaging. Bioelectrochemistry 87:253–259. doi: 10.1016/j.bioelechem.2012.03.008 CrossRefPubMedGoogle Scholar
  11. Méléard P, Bagatolli LA, Pott T (2009) Giant unilamellar vesicle electroformation from lipid mixtures to native membranes under physiological conditions. Methods Enzymol 465:161–176. doi: 10.1016/S0076-6879(09)65009-6 CrossRefPubMedGoogle Scholar
  12. Pott T, Bouvrais H, Méléard P (2008) Giant unilamellar vesicle formation under physiologically relevant conditions. Chem Phys Lipids 154:115–119. doi: 10.1016/j.chemphyslip.2008.03.008 CrossRefPubMedGoogle Scholar
  13. Reeves JP, Dowben RM (1969) Formation and properties of thin-walled phospholipid vesicles. J Cell Physiol 73:49–60. doi: 10.1002/jcp.1040730108 CrossRefPubMedGoogle Scholar
  14. Reis A, Spickett CM (2012) Chemistry of phospholipid oxidation. Biochim Biophys Acta 1818:2374–2387. doi: 10.1016/j.bbamem.2012.02.002 CrossRefPubMedGoogle Scholar
  15. Salomone F, Breton M, Leray I et al (2014) High-yield nontoxic gene transfer through conjugation of the CM18-Tat11 chimeric peptide with nanosecond electric pulses. Mol Pharm 11:2466–2474. doi: 10.1021/mp500223t CrossRefPubMedGoogle Scholar
  16. Soto-Arriaza MA, Sotomayor CP, Lissi EA (2008) Relationship between lipid peroxidation and rigidity in L-alpha-phosphatidylcholine-DPPC vesicles. J Colloid Interface Sci 323:70–74. doi: 10.1016/j.jcis.2008.04.034 CrossRefPubMedGoogle Scholar
  17. Volinsky R, Kinnunen PKJ (2013) Oxidized phosphatidylcholines in membrane-level cellular signaling: from biophysics to physiology and molecular pathology. FEBS J 280:2806–2816. doi: 10.1111/febs.12247 CrossRefPubMedGoogle Scholar
  18. Walde P, Cosentino K, Engel H, Stano P (2010) Giant vesicles: preparations and applications. Chembiochem 11:848–865. doi: 10.1002/cbic.201000010 CrossRefPubMedGoogle Scholar
  19. Wong-Ekkabut J, Xu ZT, Triampo W et al (2007) Effect of lipid peroxidation on the properties of lipid bilayers: a molecular dynamics study. Biophys J 93:4225–4236. doi: 10.1529/biophysj.107.112565 PubMedCentralCrossRefPubMedGoogle Scholar
  20. Yamamoto K, Ando J (2013) Endothelial cell and model membranes respond to shear stress by rapidly decreasing the order of their lipid phases. J Cell Sci 126:1227–1234. doi: 10.1242/jcs.119628 CrossRefPubMedGoogle Scholar
  21. Zhou Y, Berry CK, Storer PA, Raphael RM (2007) Peroxidation of polyunsaturated phosphatidyl-choline lipids during electroformation. Biomaterials 28:1298–1306. doi: 10.1016/j.biomaterials.2006.10.016 CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Marie Breton
    • 1
    • 2
    • 3
  • Mooud Amirkavei
    • 1
    • 2
    • 3
  • Lluis M. Mir
    • 1
    • 2
    • 3
  1. 1.Université Paris-Sud, Laboratoire de Vectorologie et Thérapeutiques Anticancéreuses, UMR8203OrsayFrance
  2. 2.CNRS, Laboratoire de Vectorologie et Thérapeutiques Anticancéreuses, UMR 8203OrsayFrance
  3. 3.Gustave Roussy, Laboratoire de Vectorologie et Thérapeutiques Anticancéreuses, UMR8203VillejuifFrance

Personalised recommendations