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The nucleus pulposus (NP) includes a heterogeneous cell population, which is still poorly characterized. Here, we aimed to uncover main alterations in NP cells with aging. For that, bovine coccygeal discs from young (12\u2009months) and old (10\u201316\u2009years old) animals were dissected and primary NP cells were isolated. Gene expression and proteomics of fresh NP cells were performed. NP cells were labelled with propidium iodide and analysed by flow cytometry for the expression of CD29, CD44, CD45, CD146, GD2, Tie2, CD34 and Stro\u20101. Morphological cell features were also dissected by imaging flow cytometry. Elder NP cells (up\u2010regulated bIL\u20106 and bMMP1 gene expression) presented lower percentages of CD29+, CD44+, CD45+ and Tie2+ cells compared with young NP cells (upregulated bIL\u20108, bCOL2A1 and bACAN gene expression), while GD2, CD146, Stro\u20101 and CD34 expression were maintained with age. NP cellulome showed an upregulation of proteins related to endoplasmic reticulum (ER) and melanosome independently of age, whereas proteins upregulated in elder NP cells were also associated with glycosylation and disulfide bonds. Flow cytometry analysis of NP cells disclosed the existence of 4 subpopulations with distinct auto\u2010fluorescence and size with different dynamics along aging. Regarding cell morphology, aging increases NP cell area, diameter and vesicles. These results contribute to a better understanding of NP cells aging and highlighting potential anti\u2010aging targets that can help to mitigate age\u2010related disc disease.<\/jats:p>","DOI":"10.1111\/acel.13873","type":"journal-article","created":{"date-parts":[[2023,5,31]],"date-time":"2023-05-31T05:42:53Z","timestamp":1685511773000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Alterations of bovine nucleus pulposus cells with aging"],"prefix":"10.1111","volume":"22","author":[{"given":"Maria","family":"Molinos","sequence":"first","affiliation":[{"name":"i3S \u2013 Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade Universidade do Porto  Porto Portugal"},{"name":"INEB \u2013 Instituto de Engenharia Biom\u00e9dica Universidade do Porto  Porto Portugal"},{"name":"ICBAS \u2013 Instituto de Ci\u00eancias Biom\u00e9dicas Abel Salazar Universidade do Porto  Porto 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R.","family":"Almeida","sequence":"additional","affiliation":[{"name":"i3S \u2013 Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade Universidade do Porto  Porto Portugal"},{"name":"INEB \u2013 Instituto de Engenharia Biom\u00e9dica Universidade do Porto  Porto Portugal"},{"name":"iBiMED \u2013 Institute of Biomedicine, Department of Medical Sciences University of Aveiro  Aveiro Portugal"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M\u00e1rio A.","family":"Barbosa","sequence":"additional","affiliation":[{"name":"i3S \u2013 Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade Universidade do Porto  Porto Portugal"},{"name":"INEB \u2013 Instituto de Engenharia Biom\u00e9dica Universidade do Porto  Porto Portugal"},{"name":"ICBAS \u2013 Instituto de Ci\u00eancias Biom\u00e9dicas Abel Salazar Universidade do Porto  Porto 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