Skip to main content
SpringerLink
Log in

General Protocol for the Culture of Cells on Plasma-Coated Electrospun Scaffolds

  • Protocol
  • First Online:
Stem Cell Nanotechnology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1058))

Abstract

As opposed to culture on standard tissue-treated plastic, cell culture on three-dimensional scaffolds impedes additional challenges with respect to substrate preparation, cell seeding, culture maintenance, and analysis. We herewith present a general route for the culture of primary cells, differentiated cells, or stem cells on plasma-coated, electrospun scaffolds. We describe a method to prepare and fix the scaffolds in culture wells and discuss a convenient method for cell seeding and subsequent analysis by scanning electron microscopy or immunohistology.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
eBook
USD 39.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 54.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 54.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Woodruff MA, Hutmacher DW (2010) The return of a forgotten polymer-polycaprolactone in the 21st century. Prog Polym Sci 35(10):1217–1256. doi:10.1016/j.progpolymsci.2010.04.002

    Article  CAS  Google Scholar 

  2. Cipitria A, Skelton A, Dargaville TR, Dalton PD, Hutmacher DW (2011) Design, fabrication and characterization of pcl electrospun scaffolds-a review. J Mater Chem 21(26):9419–9453. doi:10.1039/c0jm04502k

    Article  CAS  Google Scholar 

  3. Mikos AG, Thorsen AJ, Czerwonka LA, Bao Y, Langer R, Winslow DN, Vacanti JP (1994) Preparation and characterization of poly(l-lactic acid) foams. Polymer 35(5):1068–1077. doi:10.1016/0032-3861(94)90953-9

    Article  CAS  Google Scholar 

  4. Estelles JM, Vidaurre A, Duenas JMM, Cortazar IC (2008) Physical characterization of polycaprolactone scaffolds. J Mater Sci Mater Med 19(1):189–195. doi:10.1007/s10856-006-0101-2

    Article  Google Scholar 

  5. Luciani A, Coccoli V, Orsi S, Ambrosio L, Netti PA (2008) Pcl microspheres based functional scaffolds by bottom-up approach with predefined microstructural properties and release profiles. Biomaterials 29(36):4800–4807. doi:10.1016/j.biomaterials.2008.09.007

    Article  PubMed  CAS  Google Scholar 

  6. Marrazzo C, Di Maio E, Iannace S (2008) Conventional and nanometric nucleating agents in poly(epsilon-caprolactone) foaming: crystals vs. bubbles nucleation. Polym Eng Sci 48(2):336–344. doi:10.1002/pen.20937

    Article  CAS  Google Scholar 

  7. Baker SC, Atkin N, Gunning PA, Granville N, Wilson K, Wilson D, Southgate J (2006) Characterisation of electrospun polystyrene scaffolds for three-dimensional in vitro biological studies. Biomaterials 27(16):3136–3146

    Article  PubMed  CAS  Google Scholar 

  8. Formhals A (1934) A process and apparatus for preparing artificial treads. US Patent.

    Google Scholar 

  9. Woo KM, Chen VJ, Ma PX (2003) Nano-fibrous scaffolding architecture selectively enhances protein adsorption contributing to cell attachment. J Biomed Mater Res A 67A(2):531–537. doi:10.1002/jbm.a.10098

    Article  CAS  Google Scholar 

  10. Huang ZM, Zhang YZ, Kotaki M, Ramakrishna S (2003) A review on polymer nanofibers by electrospinning and their applications in nanocomposites. Compos Sci Technol 63(15):2223–2253. doi:10.1016/s0266-3538(03)00178-7

    Article  CAS  Google Scholar 

  11. Greiner A, Wendorff JH (2007) Electrospinning: a fascinating method for the preparation of ultrathin fibers. Angew Chem Int Ed 46:5670–5703

    Article  CAS  Google Scholar 

  12. Cui WG, Zhou Y, Chang J (2010) Electrospun nanofibrous materials for tissue engineering and drug delivery. Sci Technol Adv Mater 11:014108. doi:01410810.1088/1468-6996/11/1/014108

    Article  Google Scholar 

  13. Boland ED, Pawlowski KJ, Barnes CP, Simpson DG, Wnek GE, Bowlin GL (2006) Electrospinning of bioresorbable polymers for tissue engineering scaffolds. In: Reneker DH and Fong H (eds.), Polymeric nanofibers, vol 918. Acs symposium series, Oxford University Press, New York, pp 188–204.

    Google Scholar 

  14. Boudriot U, Dersch R, Greiner A, Wendorff JH (2006) Electrospinning approaches toward scaffold engineering - a brief overview. Artif Organs 30(10):785–792

    Article  PubMed  CAS  Google Scholar 

  15. Agarwal S, Greiner A, Wendorff JH (2009) Electrospinning of manmade and biopolymer nanofibers-progress in techniques, materials, and applications. Adv Funct Mater 19(18):2863–2879. doi:10.1002/adfm.200900591

    Article  CAS  Google Scholar 

  16. Curtis ASG, Forrester JV, Clark P (1986) Substrate hydroxylation and cell-adhesion. J Cell Sci 86:9–24

    CAS  Google Scholar 

  17. Guimond S, Schuetz U, Hanselmann B, Koerner E, Hegemann D (2011) Influence of gas phase and surface reactions on plasma polymerization. Surf Coat Technol 205:S447–S450. doi:10.1016/j.surfcoat.2011.03.093

    Article  CAS  Google Scholar 

  18. Hegemann D, Koerner E, Albrecht K, Schuetz U, Guimond S (2010) Growth mechanism of oxygen-containing functional plasma polymers. Plasma Processes Polym 7(11):889–898. doi:10.1002/ppap. 200900144

    Article  CAS  Google Scholar 

  19. Hegemann D, Schütz U, Körner E (2011) Macroscopic approach to plasma polymerization using the concept of energy density. Plasma Processes Polym 8(8):689–694. doi:10.1002/ppap. 201000211

    Article  CAS  Google Scholar 

  20. Koerner E, Fortunato G, Hegemann D (2009) Influence of rf plasma reactor setup on carboxylated hydrocarbon coatings. Plasma Processes Polym 6(2):119–125. doi:10.1002/ppap. 200800102

    Article  Google Scholar 

  21. Guex AG, Kocher FM, Fortunato G, Korner E, Hegemann D, Carrel TP, Tevaearai HT, Giraud MN (2012) Fine-tuning of substrate architecture and surface chemistry promotes muscle tissue development. Acta Biomater 8(4):1481–1489

    Article  PubMed  CAS  Google Scholar 

  22. Matthews JA, Wnek GE, Simpson DG, Bowlin GL (2002) Electrospinning of collagen nanofibers. Biomacromolecules 3(2):232–238. doi:10.1021/bm015533u

    Article  PubMed  CAS  Google Scholar 

  23. Moghe AK, Hufenus R, Hudson SM, Gupta BS (2009) Effect of the addition of a fugitive salt on electrospinnability of poly(epsilon-caprolactone). Polymer 50(14):3311–3318. doi:10.1016/j.polymer.2009.04.063

    Article  CAS  Google Scholar 

  24. Jaafar IH, LeBlon CE, Wei M-T, Ou-Yang D, Coulter JP, Jedlicka SS (2011) Improving fluorescence imaging of biological cells on biomedical polymers. Acta Biomater 7(4):1588–1598. doi:10.1016/j.actbio.2010.12.007

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgement

The project has been funded by the Inselspital Bern, Switzerland; the Empa St. Gallen, Switzerland; and grant No. 122334 of the Swiss National Science Foundation. The authors are grateful to Céline Fouassier for cell culture protocol optimization, Sara Brasa for protocol optimization to dehydrate samples for SEM imaging, and Enrico Körner for plasma process development and optimization.

Author information

Authors and Affiliations

  1. Swiss Federal Laboratories of Materials Science and Technology, St. Gallen, Switzerland

    A. Géraldine Guex, Giuseppino Fortunato & Dirk Hegemann

  2. Clinic of Cardiovascular Surgery, University Hospital and University of Bern, Bern, Switzerland

    Hendrik T. Tevaearai

  3. Cardiology, Department of Medicine, University of Fribourg, Fribourg, Switzerland

    Marie-Noëlle Giraud

Authors
  1. A. Géraldine Guex
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Giuseppino Fortunato
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Dirk Hegemann
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hendrik T. Tevaearai
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Marie-Noëlle Giraud
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Regenerative Medicine Program, Ottawa Hospital Research Institute Sprott Centre for Stem Cell Research, Ottawa, Canada

    Kursad Turksen

Rights and permissions

Reprints and permissions

Copyright information

© 2013 Springer Science+Business Media New York

About this protocol

Cite this protocol

Guex, A.G., Fortunato, G., Hegemann, D., Tevaearai, H.T., Giraud, MN. (2013). General Protocol for the Culture of Cells on Plasma-Coated Electrospun Scaffolds. In: Turksen, K. (eds) Stem Cell Nanotechnology. Methods in Molecular Biology, vol 1058. Humana Press, Totowa, NJ. https://doi.org/10.1007/7651_2013_8

Download citation

  • DOI: https://doi.org/10.1007/7651_2013_8

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-570-5

  • Online ISBN: 978-1-62703-571-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation