{"doi":"10.1016/j.xpro.2025.104178","title":"Protocol for the utilization of murine interstitial fluid extract to model breast cancer in the context of old age in vitro","abstract":"Interstitial fluid reflects both the organ microenvironment and tumor milieu; however, its direct study in vivo remains technically challenging. Here, we present a protocol for using murine interstitial fluid extract from lungs of metastasis-bearing mice as a model to recapitulate the metastatic environment ex vivo . This protocol outlines steps for cell preparation, tail vein injection, ex vivo tissue processing, and incubation, followed by detailed procedures for interstitial fluid isolation, in vitro cell treatment, and RNA sequencing (RNA-seq) analysis. • Instructions for intravenous injection of single-cell suspension via mouse tail vein • Guidance on ex vivo tissue processing and IFE isolation • Procedures for in vitro use of IFE followed by high-throughput screening Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Interstitial fluid reflects both the organ microenvironment and tumor milieu; however, its direct study in vivo remains technically challenging. Here, we present a protocol for using murine interstitial fluid extract from lungs of metastasis-bearing mice as a model to recapitulate the metastatic environment ex vivo . This protocol outlines steps for cell preparation, tail vein injection, ex vivo tissue processing, and incubation, followed by detailed procedures for interstitial fluid isolation, in vitro cell treatment, and RNA sequencing (RNA-seq) analysis.","journal":"STAR Protocols","year":2025,"id":579750,"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":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9422,"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":1490665,"name":"Rojan Chimeh Rad","orcid":null,"position":1,"is_corresponding":false},{"id":574892,"name":"Didem Ilter","orcid":"0000-0001-7506-8198","position":2,"is_corresponding":false},{"id":426224,"name":"Ana P. Gomes","orcid":"0000-0003-0881-727X","position":3,"is_corresponding":false},{"id":848334,"name":"Stanislav Drápela","orcid":"0000-0002-9421-9972","position":0,"is_corresponding":true}],"reference_count":11,"raw_metadata":null,"created_at":"2026-07-19T02:58:34.718602Z","pmid":"41171756","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":[]}