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Generation of Infectious Prions and Detection with the Prion-Infected Cell Assay

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Prions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1658))

Abstract

Cell lines propagating prions are an efficient and useful means for studying the cellular and molecular mechanisms implicated in prion disease. Utilization of cell-based models has led to the finding that PrPC and PrPSc are released from cells in association with extracellular vesicles known as exosomes. Exosomes have been shown to act as vehicles for infectivity, transferring infectivity between cell lines and providing a mechanism for prion spread between tissues. Here, we describe the methods for generating a prion-propagating cell line with prion-infected brain homogenate, cell lysate, conditioned media, and exosomes and also detection of protease-resistant PrP with the prion-infected cell assay.

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Author information

Authors and Affiliations

  1. The Florey Institute of Neuroscience and Mental Health, University of Melbourne, Parkville, VIC, Australia

    Laura J. Vella

  2. Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, VIC, Australia

    Bradley Coleman & Andrew F. Hill

  3. Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Bundoora, VIC, 3086, Australia

    Andrew F. Hill

Authors
  1. Laura J. Vella
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  2. Bradley Coleman
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  3. Andrew F. Hill
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Corresponding author

Correspondence to Andrew F. Hill .

Editor information

Editors and Affiliations

  1. Department of Pathology, The University of Melbourne, Parkville, Victoria, Australia

    Victoria A. Lawson

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Vella, L.J., Coleman, B., Hill, A.F. (2017). Generation of Infectious Prions and Detection with the Prion-Infected Cell Assay. In: Lawson, V. (eds) Prions. Methods in Molecular Biology, vol 1658. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7244-9_9

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  • DOI: https://doi.org/10.1007/978-1-4939-7244-9_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7242-5

  • Online ISBN: 978-1-4939-7244-9

  • eBook Packages: Springer Protocols

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