{"doi":"10.1016/j.xpro.2025.103778","title":"Protocol for the isolation and characterization of murine hematopoietic stem and progenitor cell-derived extracellular vesicles","abstract":"Hematopoietic stem cells (HSCs) maintain their self-renewal capacity in an autocrine manner through hematopoietic stem and progenitor cell (HSPC)-derived extracellular vesicles (EVs). Here, we present a protocol for the isolation and characterization of EVs from HSPCs starting from an in vivo murine model. We describe steps for murine bone marrow isolation, HSPC staining and sorting, HSPC-derived EV isolation, size and concentration characterization, and EV visualization and marker description. For complete details on the use and execution of this protocol, please refer to Bonora et al. 1 • Steps for investigating HSPC-derived EV surface markers using flow cytometry • Instructions for bone marrow HSPC isolation, staining, and sorting • Guidance on techniques for determining the size and concentration of HSPC-derived EVs • Procedures for characterizing HSPC-derived EVs Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Hematopoietic stem cells (HSCs) maintain their self-renewal capacity in an autocrine manner through hematopoietic stem and progenitor cell (HSPC)-derived extracellular vesicles (EVs). Here, we present a protocol for the isolation and characterization of EVs from HSPCs starting from an in vivo murine model. We describe steps for murine bone marrow isolation, HSPC staining and sorting, HSPC-derived EV isolation, size and concentration characterization, and EV visualization and marker description.","journal":"STAR Protocols","year":2025,"id":528127,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":2,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9506,"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":1204323,"name":"Ilaria Zanolla","orcid":"0000-0002-7395-787X","position":1,"is_corresponding":false},{"id":692340,"name":"Massimo Bonora","orcid":"0000-0002-5752-3318","position":2,"is_corresponding":false},{"id":682007,"name":"Claudia Morganti","orcid":"0000-0003-0527-4505","position":3,"is_corresponding":false},{"id":1204326,"name":"Barbara Zavan","orcid":"0000-0002-4779-4456","position":4,"is_corresponding":false},{"id":1405523,"name":"Letizia Ferroni","orcid":"0000-0002-5457-0338","position":5,"is_corresponding":false},{"id":548760,"name":"Paolo Pinton","orcid":"0000-0001-7108-6508","position":6,"is_corresponding":false},{"id":649289,"name":"Keisuke Ito","orcid":"0000-0002-6702-9735","position":7,"is_corresponding":false},{"id":1405522,"name":"Federica Zanotti","orcid":"0000-0002-8721-8162","position":0,"is_corresponding":true}],"reference_count":15,"raw_metadata":null,"created_at":"2026-07-19T02:50:44.062153Z","pmid":"40252223","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":[]}