Skip to main content
SpringerLink
Log in

Isolation and Culture of Schwann Cells

  • Protocol
  • First Online:
Neural Development

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1018))

Abstract

Primarily cultured Schwann cells are essential for the investigation of molecular mechanisms regulating proliferation, survival, differentiation, and myelination of Schwann cell and for the development of efficient transplantation for regeneration of injured spinal cord or peripheral nervous system. Here we describe a basic protocol for isolation and purification of primary Schwann cell from neonatal rat or mouse and discuss some modifications adapted to the culturing from adult nerves and optional methods for Schwann cell enrichment.

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
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. Salzer JL, Bunge RP (1980) Studies of Schwann cell proliferation. I. An analysis in tissue culture of proliferation during development, Wallerian degeneration, and direct injury. J Cell Biol 84:739–752

    Article  PubMed  CAS  Google Scholar 

  2. Jessen KR, Brennan A, Morgan L, Mirsky R, Kent A, Hashimoto Y, Gavrilovic J (1994) The Schwann cell precursor and its fate: a study of cell death and differentiation during gliogenesis in rat embryonic nerves. Neuron 12:509–527

    Article  PubMed  CAS  Google Scholar 

  3. Brockes JP, Fields KL, Raff MC (1979) Studies on cultured rat Schwann cells. I. Establishment of purified populations from cultures of peripheral nerve. Brain Res 165:105–118

    Article  PubMed  CAS  Google Scholar 

  4. Bottenstein JE, Sato GH (1979) Growth of a rat neuroblastoma cell line in serum-free supplemented medium. Proc Natl Acad Sci USA 76:514–517

    Article  PubMed  CAS  Google Scholar 

  5. Weinstein DE, Wu R (2001) Isolation and purification of primary Schwann cells. Curr Protoc Neurosci Chapter 3: Unit 3 17

    Google Scholar 

  6. Jessen KR, Mirsky R (2005) The origin and development of glial cells in peripheral nerves. Nat Rev Neurosci 6:671–682

    Article  PubMed  CAS  Google Scholar 

  7. Eldridge CF, Bunge MB, Bunge RP, Wood PM (1987) Differentiation of axon-related Schwann cells in vitro. I. Ascorbic acid regulates basal lamina assembly and myelin formation. J Cell Biol 105:1023–1034

    Article  PubMed  CAS  Google Scholar 

  8. Moya F, Bunge MB, Bunge RP (1980) Schwann cells proliferate but fail to differentiate in defined medium. Proc Natl Acad Sci USA 77:6902–6906

    Article  PubMed  CAS  Google Scholar 

  9. Haastert K, Lipokatic E, Fischer M, Timmer M, Grothe C (2006) Differentially promoted peripheral nerve regeneration by grafted Schwann cells over-expressing different FGF-2 isoforms. Neurobiol Dis 21:138–153

    Article  PubMed  CAS  Google Scholar 

  10. Haastert K, Mauritz C, Matthies C, Grothe C (2006) Autologous adult human Schwann cells genetically modified to provide alternative cellular transplants in peripheral nerve regeneration. J Neurosurg 104:778–786

    Article  PubMed  Google Scholar 

  11. Takagi T, Ishii K, Shibata S, Yasuda A, Sato M, Nagoshi N, Saito H, Okano HJ, Toyama Y, Okano H, Nakamura M (2011) Schwann-spheres derived from injured peripheral nerves in adult mice–their in vitro characterization and therapeutic potential. PLoS One 6:e21497

    Article  PubMed  CAS  Google Scholar 

  12. Scarpini E, Kreider BQ, Lisak RP, Pleasure DE (1988) Establishment of Schwann cell cultures from adult rat peripheral nerves. Exp Neurol 102:167–176

    Article  PubMed  CAS  Google Scholar 

  13. Kim HA, Pomeroy SL, Whoriskey W, Pawlitzky I, Benowitz LI, Sicinski P, Stiles CD, Roberts TM (2000) A developmentally regulated switch directs regenerative growth of Schwann cells through cyclin D1. Neuron 26:405–416

    Article  PubMed  CAS  Google Scholar 

  14. Stevens B, Tanner S, Fields RD (1998) Control of myelination by specific patterns of neural impulses. J Neurosci 18:9303–9311

    PubMed  CAS  Google Scholar 

  15. Seilheimer B, Schachner M (1987) Regulation of neural cell adhesion molecule expression on cultured mouse Schwann cells by nerve growth factor. EMBO J 6:1611–1616

    PubMed  CAS  Google Scholar 

  16. Dong Z, Sinanan A, Parkinson D, Parmantier E, Mirsky R, Jessen KR (1999) Schwann cell development in embryonic mouse nerves. J Neurosci Res 56:334–348

    Article  PubMed  CAS  Google Scholar 

  17. Haastert K, Mauritz C, Chaturvedi S, Grothe C (2007) Human and rat adult Schwann cell cultures: fast and efficient enrichment and highly effective non-viral transfection protocol. Nat Protoc 2:99–104

    Article  PubMed  CAS  Google Scholar 

  18. Mauritz C, Grothe C, Haastert K (2004) Comparative study of cell culture and ­purification methods to obtain highly enriched cultures of proliferating adult rat Schwann cells. J Neurosci Res 77:453–461

    Article  PubMed  CAS  Google Scholar 

  19. Tao Y, Dai P, Liu Y, Marchetto S, Xiong WC, Borg JP, Mei L (2009) Erbin regulates NRG1 signaling and myelination. Proc Natl Acad Sci USA 106:9477–9482

    Article  PubMed  CAS  Google Scholar 

  20. Salzer JL, Bunge RP, Glaser L (1980) Studies of Schwann cell proliferation. III. Evidence for the surface localization of the neurite mitogen. J Cell Biol 84:767–778

    Article  PubMed  CAS  Google Scholar 

  21. Dong Z, Brennan A, Liu N, Yarden Y, Lefkowitz G, Mirsky R, Jessen KR (1995) Neu differentiation factor is a neuron-glia signal and regulates survival, proliferation, and maturation of rat Schwann cell precursors. Neuron 15:585–596

    Article  PubMed  CAS  Google Scholar 

  22. Manthorpe M, Skaper S, Varon S (1980) Purification of mouse Schwann cells using neurite-induced proliferation in serum-free monolayer culture. Brain Res 196:467–482

    Article  PubMed  CAS  Google Scholar 

  23. Salzer JL, Williams AK, Glaser L, Bunge RP (1980) Studies of Schwann cell proliferation. II. Characterization of the stimulation and specificity of the response to a neurite membrane fraction. J Cell Biol 84:753–766

    Article  PubMed  CAS  Google Scholar 

  24. Schumacher M, Jung-Testas I, Robel P, Baulieu EE (1993) Insulin-like growth factor I: a mitogen for rat Schwann cells in the presence of elevated levels of cyclic AMP. Glia 8:232–240

    Article  PubMed  CAS  Google Scholar 

  25. Taveggia C, Zanazzi G, Petrylak A, Yano H, Rosenbluth J, Einheber S, Xu X, Esper RM, Loeb JA, Shrager P, Chao MV, Falls DL, Role L, Salzer JL (2005) Neuregulin-1 type III determines the ensheathment fate of axons. Neuron 47:681–694

    Article  PubMed  CAS  Google Scholar 

  26. Haastert K, Semmler N, Wesemann M, Rucker M, Gellrich NC, Grothe C (2006) Establishment of cocultures of osteoblasts, Schwann cells, and neurons towards a tissue-engineered approach for orofacial ­reconstruction. Cell Transplant 15:733–744

    Article  PubMed  Google Scholar 

  27. Morrissey TK, Kleitman N, Bunge RP (1991) Isolation and functional characterization of Schwann cells derived from adult peripheral nerve. J Neurosci 11:2433–2442

    PubMed  CAS  Google Scholar 

  28. Komiyama T, Nakao Y, Toyama Y, Asou H, Vacanti CA, Vacanti MP (2003) A novel technique to isolate adult Schwann cells for an artificial nerve conduit. J Neurosci Methods 122:195–200

    Article  PubMed  Google Scholar 

  29. Chen ZL, Strickland S (2003) Laminin gamma1 is critical for Schwann cell differentiation, axon myelination, and regeneration in the peripheral nerve. J Cell Biol 163:889–899

    Article  PubMed  CAS  Google Scholar 

  30. Yu WM, Feltri ML, Wrabetz L, Strickland S, Chen ZL (2005) Schwann cell-specific ablation of laminin gamma1 causes apoptosis and prevents proliferation. J Neurosci 25:4463–4472

    Article  PubMed  CAS  Google Scholar 

  31. Armstrong SJ, Wiberg M, Terenghi G, Kingham PJ (2007) ECM molecules mediate both Schwann cell proliferation and activation to enhance neurite outgrowth. Tissue Eng 13:2863–2870

    Article  PubMed  CAS  Google Scholar 

  32. Needham LK, Tennekoon GI, McKhann GM (1987) Selective growth of rat Schwann cells in neuron- and serum-free primary culture. J Neurosci 7:1–9

    PubMed  CAS  Google Scholar 

  33. Morgan L, Jessen KR, Mirsky R (1991) The effects of cAMP on differentiation of cultured Schwann cells: progression from an early phenotype (04+) to a myelin phenotype (P0+, GFAP-, N-CAM-, NGF-receptor-) depends on growth inhibition. J Cell Biol 112:457–467

    Article  PubMed  CAS  Google Scholar 

  34. Mahanthappa NK, Anton ES, Matthew WD (1996) Glial growth factor 2, a soluble neuregulin, directly increases Schwann cell motility and indirectly promotes neurite outgrowth. J Neurosci 16:4673–4683

    PubMed  CAS  Google Scholar 

  35. Haynes LW, Rushton JA, Perrins MF, Dyer JK, Jones R, Howell R (1994) Diploid and hyperdiploid rat Schwann cell strains displaying negative autoregulation of growth in vitro and myelin sheath-formation in vivo. J Neurosci Methods 52:119–127

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Development and Regenerative Biology, Hangzhou Normal University, Hangzhou, Zhejiang, China

    Yanmei Tao

Authors
  1. Yanmei Tao
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. , Department of Chemical Biology, Rutgers University, Frelinghuysen Road 164, Piscataway, 08854, New Jersey, USA

    Renping Zhou

  2. , Inst. Molecular Medicine & Genetics, Georgia Health Sciences University, 15th Street 1120, Augusta, 30912, Georgia, USA

    Lin Mei

Rights and permissions

Reprints and permissions

Copyright information

© 2013 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Tao, Y. (2013). Isolation and Culture of Schwann Cells. In: Zhou, R., Mei, L. (eds) Neural Development. Methods in Molecular Biology, vol 1018. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-444-9_9

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-444-9_9

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-443-2

  • Online ISBN: 978-1-62703-444-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation