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Design of a Versatile Sample Holder for Facile Culture of Cells on Electrospun Membranes or Thin Polymer Films Under Flow Conditions

  • Anne-Sophie Mertgen
  • Markus Rottmar
  • Lukas Weidenbacher
  • Anne Géraldine GuexEmail author
Protocol
Part of the Methods in Molecular Biology book series

Abstract

Endothelial cell culture under flow, to mimic physiological conditions within blood vessels, has gained particular attention for the formation of a homogeneous endothelium in vitro. Here, we report on the design of a setup for simultaneous culture of up to nine electrospun membranes or thin polymer films in custom-made holders under flow on an orbital shaker. The versatile design of the device allows for the use of electrospun membranes/polymer films of choice and subsequent analysis with commonly used methods such as immunofluorescence or scanning electron microscopy.

Keywords

Culture under flow Electrospun membranes Endothelial cells 

Notes

Acknowledgments

We would like to thank Jörg Gschwend, Empa, St. Gallen for material manufacture and his assistance for device design. We are also grateful to Marielle Hintereder, Empa, St. Gallen, for assistance during cell culture. Furthermore, we would like to acknowledge the input from Andreia Fernandes and Seyedvahid Hosseini, Laboratory for Applied Mechanobiology, ETH Zürich for setting up cell culture on an orbital shaker. This work is part of the Zurich Heart project under the umbrella of University Medicine Zurich/Hochschulmedizin Zürich.

References

  1. 1.
    Levesque MJ, Nerem RM (1985) The elongation and orientation of cultured endothelial cells in response to shear stress. J Biomech Eng 107(4):341–347Google Scholar
  2. 2.
    Schnittler HJ, Franke RP, Akbay U, Mrowietz C, Drenckhahn D (1993) Improved in vitro rheological system for studying the effect of fluid shear stress on cultured cells. Am J Phys 265(1 Pt 1):C289–C298Google Scholar
  3. 3.
    Whited BM, Rylander MN (2014) The influence of electrospun scaffold topography on endothelial cell morphology, alignment, and adhesion in response to fluid flow. Biotechnol Bioeng 111(1):184–195Google Scholar
  4. 4.
    Gong X, Liu H, Ding X, Liu M, Li X, Zheng L, Jia X, Zhou G, Zou Y, Li J, Huang X, Fan Y (2014) Physiological pulsatile flow culture conditions to generate functional endothelium on a sulfated silk fibroin nanofibrous scaffold. Biomaterials 35(17):4782–4791Google Scholar
  5. 5.
    Dardik A, Chen L, Frattini J, Asada H, Aziz F, Kudo FA, Sumpio BE (2005) Differential effects of orbital and laminar shear stress on endothelial cells. J Vasc Surg 41(5):869–880Google Scholar
  6. 6.
    Salek MM, Sattari P, Martinuzzi RJ (2012) Analysis of fluid flow and wall shear stress patterns inside partially filled agitated culture well plates. Ann Biomed Eng 40(3):707–728Google Scholar
  7. 7.
    Filipovic N, Ghimire K, Saveljic I, Milosevic Z, Ruegg C (2016) Computational modeling of shear forces and experimental validation of endothelial cell responses in an orbital well shaker system. Comput Methods Biomech Biomed Engin 19(6):581–590Google Scholar
  8. 8.
    Guex AG, Weidenbacher L, Maniura-Weber K, Rossi RM, Fortunato G (2017) Hierarchical self-assembly of poly(urethane)/poly(vinylidene fluoride-co-hexafluoropropylene) blends into highly hydrophobic electrospun fibers with reduced protein adsorption profiles. Macromol Mater Eng 302(10):1700081Google Scholar
  9. 9.
    Papaioannou TG, Karatzis EN, Vavuranakis M, Lekakis JP, Stefanadis C (2006) Assessment of vascular wall shear stress and implications for atherosclerotic disease. Int J Cardiol 113(1):12–18Google Scholar
  10. 10.
    Guex AG, Fortunato G, Hegemann D, Tevaearai HT, Giraud MN (2013) General protocol for the culture of cells on plasma-coated electrospun scaffolds. Methods Mol Biol 1058:119–131Google Scholar
  11. 11.
    Oliveira VC, Carrara RC, Simoes DL, Saggioro FP, Carlotti CG Jr, Covas DT, Neder L (2010) Sudan Black B treatment reduces autofluorescence and improves resolution of in situ hybridization specific fluorescent signals of brain sections. Histol Histopathol 25(8):1017–1024Google Scholar

Copyright information

© Springer Science+Business Media New York 2018

Authors and Affiliations

  • Anne-Sophie Mertgen
    • 1
    • 2
  • Markus Rottmar
    • 1
  • Lukas Weidenbacher
    • 1
    • 2
  • Anne Géraldine Guex
    • 1
    • 2
    Email author
  1. 1.Empa, Swiss Federal Laboratories for Materials Science and TechnologyLaboratory for BiointerfacesSt. GallenSwitzerland
  2. 2.Empa, Swiss Federal Laboratories for Materials Science and TechnologyLaboratory for Biomimetic Membranes and TextilesSt. GallenSwitzerland

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