Abstract
Methods enabling prion replication ex vivo are important for advancing prion science. However, few such technologies exist and many prion strains are intractable with them. Here, we describe a prion organotypic slice culture assay (POSCA), which allows for prion amplification and titration ex vivo under conditions that closely resemble intracerebral infection. Organotypic slices are incubated with infectious inoculum as free-floating sections, washed and cultured for up to 8 weeks. Slice cultures are a rich source of protein or RNA and allow for stringent comparisons between uninfected and prion-infected samples generated from the same mouse. Thirty-five days after contact with prions, cerebellar slices have amplified PrPSc quantitatively similar to that seen in vivo, but accelerated fivefold. The POSCA detects replication of specific prion strains from disparate sources, including bovines and ovines, with variable efficiency. The culture procedure and prion infection can be performed in 8 h.
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Acknowledgements
We thank Lotty Rietschin and Beat Gähwiler for help with slice cultures; Denis Marino, Audrey Marcel, Christian Julius, Ilan Margalith, Petra Schwarz and Frank L. Heppner for technical help. A.A. is supported by grants of the EU (TSEUR), the Swiss National Foundation, the National Competence Center on Neural Plasticity and Repair, the Stammbach foundation and DEFRA. J.F. is supported by a grant from Zentrum für Neurowissenschaften Zurich (ZNZ), the Desiree and Niels Yde foundation, the Swiss center of transgenic expertise, the Henny Sophie Clausen og möbelarkitekt Axel Clausens Foundation and the Ivan Nielsens Foundation.
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Falsig, J., Aguzzi, A. The prion organotypic slice culture assay—POSCA. Nat Protoc 3, 555–562 (2008). https://doi.org/10.1038/nprot.2008.13
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DOI: https://doi.org/10.1038/nprot.2008.13
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