Advertisement

A Self-Assembled Antifouling Nano-Biointerface for the Generation of Spheroids

  • Christoph Eilenberger
  • Mario Rothbauer
  • Peter Ertl
  • Seta Küpcü
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1771)

Abstract

Several techniques have been established over the last decades to produce three-dimensional (3D) cellular spheroids and each method has its advantages and limitations. The unique self-assembly properties of surface layer (S-layer) proteins have already been applied to a broad range of life science applications. The bacterial S-layer protein SbpA displays a strong antifouling behavior when recrystallized on planar surfaces and offers the opportunity to induce 3D cell aggregation. In this chapter, an S-layer nanointerface is presented as novel ultralow attachment material for the formation of functional spheroids of reproducible sizes. The system is compatible with standard microwell plates and enables long-term 3D cell culture and in situ monitoring of cellular viability. Moreover, this facile and stable biointerface has potential for use in toxicity screening assays and represents an alternative to conventional materials like polyethylene glycol (PEG), agarose, or hydrogel surfaces.

Key words

3D cell culture Spheroids Nanointerface S-layer Live-Dead assay TEM 

References

  1. 1.
    Breslin S, O’Driscoll L (2013) Three-dimensional cell culture: the missing link in drug discovery. Drug Discov Today 18:240–249CrossRefGoogle Scholar
  2. 2.
    Greek R, Menache A (2013) Systematic reviews of animal models: methodology versus epistemology. Int J Med Sci 10:206–221CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Zhang C, Zhao Z, Abdul Rahim N et al (2009) Towards a human-on-chip: culturing multiple cell types on a chip with compartmentalized microenvironments. Lab Chip 9:3185–3192CrossRefGoogle Scholar
  4. 4.
    Mehta G, Hsiao A, Ingram M et al (2012) Opportunities and challenges for use of tumor spheroids as models to test drug delivery and efficacy. J Control Release 164:192–204CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Dang SM, Kyba M, Perlingeiro R et al (2002) Efficiency of embryoid body formation and hematopoietic development from embryonic stem cells in different culture systems. Biotechnol Bioeng 78:442–453CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Koike M, Kurosawa H, Amano Y (2005) A round-bottom 96-well polystyrene plate coated with 2-methacryloyloxyethyl phosphorylcholine as an effective tool for embryoid body formation. Cytotechnology 47:3–10CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Corning® Ultra-Low Attachment Products (2007) Corning Inc. http://www.corning.com/lifesciences. Accessed 22 Feb 2017
  8. 8.
    Ilk N, Völlenkle C, Egelseer E et al (2002) Molecular characterization of the S-layer gene, sbpA, of Bacillus sphaericus CCM 2177 and production of a functional S-layer fusion protein with the ability to recrystallize in a defined orientation while presenting the fused allergen. Appl Environ Microbiol 68:3251–3260CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Moreno-Flores S, Küpcü S (2015) 2D protein arrays induce 3D in vivo-like assemblies of cells. Soft Matter 11:1259–1264CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Christoph Eilenberger
    • 1
    • 2
    • 3
  • Mario Rothbauer
    • 2
    • 3
  • Peter Ertl
    • 2
    • 3
  • Seta Küpcü
    • 1
  1. 1.Department of Nanobiotechnology, Institute of Synthetic BioarchitecturesUniversity of Natural Resources and Life Sciences, ViennaViennaAustria
  2. 2.Faculty of Technical Chemistry, Institute of Applied Synthetic ChemistryVienna University of TechnologyViennaAustria
  3. 3.Faculty of Technical Chemistry, Institute of Chemical Technologies and Analytics (CTA)Vienna University of TechnologyViennaAustria

Personalised recommendations