{"doi":"10.1016/j.xpro.2025.103787","title":"Protocol for assessing antibody-dependent cellular phagocytosis by primary murine and human macrophages","abstract":"Macrophages play a crucial role in mediating antibody-dependent cellular phagocytosis (ADCP), a process that enhances the effectiveness of several cancer immunotherapies. In this protocol, we outline detailed steps for isolating, differentiating, and polarizing macrophages from both mouse and human sources. Additionally, we describe an optimized technique to assess ADCP using in vitro co-culture studies followed by flow cytometry analysis. This protocol offers a reliable approach to evaluate the efficiency of macrophage-mediated phagocytosis of cancer cells during treatment with antibody-based therapies. For complete details on the use and execution of this protocol, please refer to Osorio et al. 1 • Isolation, differentiation, and polarization of mouse BMDMs and human MDMs in vitro • Phagocytosis of live tumor cells by antibody-treated primary mouse or human macrophages • Flow cytometry analysis of antibody-dependent cellular phagocytosis of cancer cells Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Macrophages play a crucial role in mediating antibody-dependent cellular phagocytosis (ADCP), a process that enhances the effectiveness of several cancer immunotherapies. In this protocol, we outline detailed steps for isolating, differentiating, and polarizing macrophages from both mouse and human sources. Additionally, we describe an optimized technique to assess ADCP using in vitro co-culture studies followed by flow cytometry analysis. This protocol offers a reliable approach to evaluate the efficiency of macrophage-mediated phagocytosis of cancer cells during treatment with antibody-based therapies.","journal":"STAR Protocols","year":2025,"id":552507,"datarank":0.10397207708399181,"base_score":0.6931471805599453,"endowment":0.6931471805599453,"self_citation_contribution":0.10397207708399181,"citation_network_contribution":0.0,"self_endowment_contribution":0.10397207708399181,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9567,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1374729,"name":"Andrew Mijacika","orcid":"0009-0007-0464-9842","position":1,"is_corresponding":false},{"id":1448904,"name":"Bella Somoza","orcid":null,"position":2,"is_corresponding":false},{"id":864613,"name":"Juan C. Osorio","orcid":"0000-0001-8090-1551","position":3,"is_corresponding":false},{"id":1224337,"name":"Carlo Sevilla","orcid":null,"position":0,"is_corresponding":true}],"reference_count":2,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T02:54:33.203144Z","pmid":"40279242","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}