{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,2]],"date-time":"2026-03-02T17:03:19Z","timestamp":1772470999772,"version":"3.50.1"},"reference-count":24,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,2,28]],"date-time":"2026-02-28T00:00:00Z","timestamp":1772236800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Italian Ministry of Health, \u201cRicerca Corrente\u201d"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Background: Advanced therapy medicinal products require rigorous preclinical testing to exclude tumorigenicity. Human articular cartilage cells expanded at low density with human platelet lysate show enhanced proliferation, matrix production, and immunomodulatory properties, supporting their use for diffuse cartilage lesions in osteoarthritic joints. This study evaluated tumorigenicity and biodistribution of cartilage cell spheroids generated using two platelet lysate sources. Methods: Cartilage cells were expanded at low density with two platelet lysates and assembled into spheroids. Cytogenetic stability was assessed by metaphase karyotyping following expansion. Immunodeficient mice received subcutaneous implantation and were monitored for 180 days. Human colon carcinoma cells and mouse fibroblasts were used as controls. Clinical follow-up, full organ histopathology, and immunohistochemistry were performed to detect human cell persistence. Results: Expanded cartilage cells showed predominantly normal karyotypes, with rare low-level mosaic chromosomal alterations not detected at the previous passage. Cartilage cell spheroids were well tolerated in vivo, with complete survival and no evidence of tumorigenicity, inflammation, or human cell persistence at implantation sites or distant organs. Control experiments confirmed the sensitivity of the model, and no systemic toxicity was observed. Conclusions: Spheroids derived from cartilage cells are non-tumorigenic, non-migratory, and biologically safe in immunodeficient mice. These findings support their development as cell-based cartilage therapies and align with regulatory recommendations for non-clinical safety evaluation.<\/jats:p>","DOI":"10.3390\/cells15050429","type":"journal-article","created":{"date-parts":[[2026,3,2]],"date-time":"2026-03-02T16:06:59Z","timestamp":1772467619000},"page":"429","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Preclinical Tumorigenicity Study of an Advanced Therapy Medicinal Product for Diffuse Cartilage Lesions in an Osteoarthritic Environment"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1800-3424","authenticated-orcid":false,"given":"Alessandra","family":"Colombini","sequence":"first","affiliation":[{"name":"Orthopaedic Biotechnology Lab, IRCCS Galeazzi Sant\u2019Ambrogio Hospital, 20157 Milan, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-4658-722X","authenticated-orcid":false,"given":"Vincenzo","family":"Raffo","sequence":"additional","affiliation":[{"name":"Department of Precision and Regenerative Medicine and Ionian Area (Di.Me.Pre-J), University of Bari, 70124 Bari, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8752-3231","authenticated-orcid":false,"given":"Vincenzo","family":"Pennone","sequence":"additional","affiliation":[{"name":"Cell and Tissue Engineering Laboratory, IRCCS Galeazzi Sant\u2019Ambrogio Hospital, 20157 Milan, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2889-0394","authenticated-orcid":false,"given":"Katia","family":"Mareschi","sequence":"additional","affiliation":[{"name":"Stem Cell Transplantation and Cellular Therapy Laboratory, Paediatric Onco-Haematology Division, Regina Margherita Children\u2019s Hospital, City of Health and Science of Turin, 10126 Turin, Italy"},{"name":"Department of Public Health and Paediatrics, University of Turin, 10126 Turin, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luciana","family":"Labanca","sequence":"additional","affiliation":[{"name":"Blood Component Production and Validation Center, City of Health and Science of Turin, S. Anna Hospital, 10126 Turin, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Laura","family":"Mangiavini","sequence":"additional","affiliation":[{"name":"IRCCS Galeazzi Sant\u2019Ambrogio Hospital, 20157 Milan, Italy"},{"name":"Department of Biomedical Sciences for Health, University of Milan, 20133 Milan, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Matteo","family":"Moretti","sequence":"additional","affiliation":[{"name":"Cell and Tissue Engineering Laboratory, IRCCS Galeazzi Sant\u2019Ambrogio Hospital, 20157 Milan, Italy"},{"name":"Regenerative Medicine Division, Institute for Translational Research, Universit\u00e0 della Svizzera Italiana (USI)\u2013Ente Ospedaliero Cantonale (EOC), 6500 Bellinzona, Switzerland"},{"name":"Service of Orthopaedics and Traumatology, Department of Surgery, EOC, 6900 Lugano, Switzerland"},{"name":"Euler Institute, Faculty of Biomedical Sciences, Universit\u00e0 della Svizzera Italiana (USI), 6900 Lugano, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7172-9662","authenticated-orcid":false,"given":"Camilla","family":"Recordati","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine and Animal Science, University of Milan, 26900 Lodi, Italy"},{"name":"Mouse and Animal Pathology Laboratory, Fondazione UNIMI, 20139 Milan, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2578-4409","authenticated-orcid":false,"given":"Federico","family":"Armando","sequence":"additional","affiliation":[{"name":"Department of Veterinary Sciences, University of Parma, 43126 Parma, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9979-3092","authenticated-orcid":false,"given":"Laura de","family":"Girolamo","sequence":"additional","affiliation":[{"name":"Orthopaedic Biotechnology Lab, IRCCS Galeazzi Sant\u2019Ambrogio Hospital, 20157 Milan, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5230-9718","authenticated-orcid":false,"given":"Arianna B.","family":"Lovati","sequence":"additional","affiliation":[{"name":"Cell and Tissue Engineering Laboratory, IRCCS Galeazzi Sant\u2019Ambrogio Hospital, 20157 Milan, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,28]]},"reference":[{"key":"ref_1","unstructured":"European Medicines Agency (EMA) (2026, January 27). 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