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Mesenchymal stromal cells (MSCs) show promise in TBI treatment through their secretome (conditioned media, CM). A low\u2010molecular\u2010weight (&lt;700\u00a0Da) CM fraction with neuroprotective effects comparable to total CM after acute brain injury in vitro is previously identified. Here, it is aimed at identifying key bioactive factors, reconstituting them into a synthetic cocktail (SYNT), and evaluating its efficacy in TBI models. Metabolomic profiling identified three prostaglandins and kynurenine, which are used to create SYNT. The SYNT formulation reduced cell death, neuronal damage, and induced protective gene expression changes associated with neuronal protection and microglia modulation toward beneficial phenotype after TBI in vitro. In vivo, SYNT conferred similar long\u2010term functional benefits as CM, improving sensorimotor function up to 6 months and memory preservation at 4 months compared to saline\u2010treated animals, though only CM reduced contusion volume at 5 months. Both treatments modulated neuroinflammation, evidenced by reduced microglial activation and astrogliosis in the pericontusional tissue at 6 months. These findings demonstrate the neuroprotective effects of MSC\u2010secretome treatment in TBI and highlight prostaglandins and kynurenine as key mediators of this response. The findings lay the groundwork for developing a standardized, cell\u2010free therapeutic strategy for TBI based on MSC derivatives.<\/jats:p>","DOI":"10.1002\/advs.202415508","type":"journal-article","created":{"date-parts":[[2025,6,19]],"date-time":"2025-06-19T06:40:21Z","timestamp":1750315221000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Mesenchymal Stromal Cell Secretome and Its Key Bioactive Metabolites Induce Long\u2010Term Neuroprotection After Traumatic Brain Injury in Mice"],"prefix":"10.1002","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7151-0812","authenticated-orcid":false,"given":"Francesca","family":"Pischiutta","sequence":"first","affiliation":[{"name":"Laboratory of Traumatic Brain Injury and Neuroprotection Department of Acute Brain and Cardiovascular Injury Istituto di Ricerche Farmacologiche Mario Negri IRCCS  Milan Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-2446-2758","authenticated-orcid":false,"given":"Francesca","family":"Tribuzio","sequence":"additional","affiliation":[{"name":"Laboratory of Traumatic Brain Injury and Neuroprotection Department of Acute Brain and Cardiovascular Injury Istituto di Ricerche Farmacologiche Mario Negri IRCCS  Milan Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7883-4679","authenticated-orcid":false,"given":"Marta","family":"Magatti","sequence":"additional","affiliation":[{"name":"Centro di Ricerca E. 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