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However, the distinct physicochemical properties of \u03b1Syn amyloid fibrils can potentially reduce their seeding activity, especially during storage. In this study, it is demonstrated that \u03b1Syn PFFs exhibit significant sensitivity to low temperatures, with notable denaturation occurring between \u221220 and 4\u00a0\u00b0C, and gradual disassembly persisted even under storage conditions at \u221280\u00a0\u00b0C. To mitigate this issue, a commonly used protein stabilizer, glycerol is introduced, which significantly reverses the cold\u2010induced disassembly of PFFs. Remarkably, storing PFFs with 20% glycerol at \u221280\u00a0\u00b0C for a month preserved their morphology and seeding activity as freshly prepared PFFs. Glycerol\u2010stabilized \u03b1Syn PFFs resulted in compromised neuronal survival, with the extent of these impairments correlating with the formation of \u03b1Syn pathology both in vivo and in vitro, indistinguishable from freshly prepared PFFs. Storing sonicated PFFs with 20% glycerol at \u221280\u00a0\u00b0C provides an optimal storage method, as sonication is necessary for activating their seeding potential. This approach reduces the frequency of sonication, simplifies handling, and ultimately lowers the overall workload, enhancing the practicality of using PFFs.<\/jats:p>","DOI":"10.1002\/smll.202401552","type":"journal-article","created":{"date-parts":[[2024,10,1]],"date-time":"2024-10-01T01:02:38Z","timestamp":1727744558000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Optimal Preservation of PFFs in Glycerol Enhances Suitability for Modeling Parkinson's Disease"],"prefix":"10.1002","volume":"20","author":[{"given":"Hengxu","family":"Mao","sequence":"first","affiliation":[{"name":"Department of Neurology the First Affiliated Hospital of Guangzhou Medical University  Guangzhou 510120 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yaoyun","family":"Kuang","sequence":"additional","affiliation":[{"name":"Department of Neurology the First Affiliated Hospital of Guangzhou Medical University  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