Vella Laura J, Coleman Bradley, Hill Andrew F
The Florey Institute of Neuroscience and Mental Health, University of Melbourne, Parkville, VIC, Australia.
Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, VIC, Australia.
Methods Mol Biol. 2017;1658:105-118. doi: 10.1007/978-1-4939-7244-9_9.
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 PrP and PrP 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.
传播朊病毒的细胞系是研究朊病毒疾病所涉及的细胞和分子机制的一种有效且有用的手段。基于细胞的模型的应用已导致发现朊蛋白（PrP）与称为外泌体的细胞外囊泡相关联地从细胞中释放出来。外泌体已被证明可作为感染性的载体，在细胞系之间传递感染性，并为朊病毒在组织间传播提供一种机制。在此，我们描述了用朊病毒感染的脑匀浆、细胞裂解物、条件培养基和外泌体生成朊病毒传播细胞系的方法，以及通过朊病毒感染细胞测定法检测蛋白酶抗性PrP的方法。
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