{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T22:14:54Z","timestamp":1790892894129,"version":"4.1.0"},"reference-count":9,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2026,4,22]],"date-time":"2026-04-22T00:00:00Z","timestamp":1776816000000},"content-version":"vor","delay-in-days":21,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2026,4,1]],"date-time":"2026-04-01T00:00:00Z","timestamp":1775001600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2026,4]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    We present a reproducible in vitro protocol for harvesting and culturing murine bone marrow\u2010derived macrophages, with the added capability of freezing bone marrow cells at \u201380\u00b0C for scalability and long\u2010term storage. To induce macrophage senescence, we developed a genotoxic stress\u2010based method using either 10 Gy ionizing radiation or 500 nM doxorubicin treatment. The resulting senescent macrophages exhibit key hallmarks of cellular senescence, including irreversible cell cycle arrest, upregulation of senescence\u2010associated markers (e.g.,\n                    <jats:italic>Cdkn1a<\/jats:italic>\n                    ), secretion of senescence\u2010associated secretory phenotype (SASP) factors, morphological changes, and SA\u2010\u03b2\u2010galactosidase activity. This model serves as a valuable tool for investigating macrophage senescence, a relatively understudied senescent cell type, and provides mechanistic insights into the contribution of the innate immune system to aging and age\u2010related diseases. \u00a9 2026 The Author(s).\n                    <jats:italic>Current Protocols<\/jats:italic>\n                    published by Wiley Periodicals LLC.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 1<\/jats:bold>\n                    : Mouse dissection and bone marrow harvest\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 2<\/jats:bold>\n                    : Thawing and plating cryopreserved murine bone marrow for macrophage differentiation\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol 1<\/jats:bold>\n                    : Flow cytometry validation of macrophage surface markers\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol 2<\/jats:bold>\n                    : Gene expression analysis via RT\u2010qPCR\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 3<\/jats:bold>\n                    : Inducing senescence in macrophages using doxorubicin or irradiation\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Alternate Protocol<\/jats:bold>\n                    : Maintenance and expansion of control macrophages\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol 3<\/jats:bold>\n                    : Brightfield microscopy and SA\u2010\u03b2\u2010galactosidase staining to assess senescence\u2010associated morphology\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol 4<\/jats:bold>\n                    : Quantifying senescence and SASP marker expression by qPCR and\/or western blot\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol 5<\/jats:bold>\n                    : Assessment of cell cycle arrest using EdU labeling with optional DNA content staining\n                  <\/jats:p>","DOI":"10.1002\/cpz1.70230","type":"journal-article","created":{"date-parts":[[2026,4,22]],"date-time":"2026-04-22T10:27:25Z","timestamp":1776853645000},"source":"Crossref","is-referenced-by-count":2,"title":["An In Vitro Model of Macrophage Senescence"],"prefix":"10.1002","volume":"6","author":[{"given":"Grasiela","family":"Torres","sequence":"first","affiliation":[{"name":"Microbiology, Immunology, Molecular Genetics University of California Los Angeles  Los Angeles California"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Utkarsh","family":"Tripathi","sequence":"additional","affiliation":[{"name":"Microbiology, Immunology, Molecular Genetics University of California Los Angeles  Los Angeles California"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ivan","family":"Salladay\u2010Perez","sequence":"additional","affiliation":[{"name":"Microbiology, Immunology, Molecular Genetics University of California Los Angeles  Los Angeles California"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Itzetl","family":"Avila","sequence":"additional","affiliation":[{"name":"Microbiology, Immunology, Molecular Genetics University of California Los Angeles  Los Angeles California"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7884-652X","authenticated-orcid":false,"given":"Anthony J.","family":"Covarrubias","sequence":"additional","affiliation":[{"name":"Microbiology, Immunology, Molecular Genetics University of California Los Angeles  Los Angeles California"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,4,22]]},"reference":[{"key":"e_1_2_14_2_1","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.202006017"},{"key":"e_1_2_14_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/9780471729259.mca02as00"},{"key":"e_1_2_14_4_1","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2022.915110"},{"key":"e_1_2_14_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41419\u2010022\u201005089\u2010y"},{"key":"e_1_2_14_6_1","doi-asserted-by":"publisher","DOI":"10.3390\/biom10030420"},{"key":"e_1_2_14_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.xpro.2023.102624"},{"key":"e_1_2_14_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/s43587-026-01101-6"},{"key":"e_1_2_14_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/cpz1.70355"},{"key":"e_1_2_14_10_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12964\u2010021\u201000718\u20104"}],"container-title":["Current Protocols"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpz1.70230","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/cpz1.70230","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpz1.70230","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,28]],"date-time":"2026-04-28T10:42:13Z","timestamp":1777372933000},"score":1,"resource":{"primary":{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/10.1002\/cpz1.70230"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4]]},"references-count":9,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2026,4]]}},"alternative-id":["10.1002\/cpz1.70230"],"URL":"https:\/\/doi.org\/10.1002\/cpz1.70230","archive":["Portico"],"relation":{},"ISSN":["2691-1299","2691-1299"],"issn-type":[{"value":"2691-1299","type":"print"},{"value":"2691-1299","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4]]},"article-number":"e70230"}}