{"doi":"10.1016/j.xpro.2025.104174","title":"Protocol for mitochondria lipid encapsulation using a dual-tube system for mitochondria transfer therapy","abstract":"Mitochondria transplantation therapy is emerging as a novel therapeutic approach to promote neuroprotection in central nervous system (CNS) disorders. Here, we present a protocol for mitochondrial surface modification that enhances the restoration of intracellular adenosine triphosphate (ATP) levels under conditions of oxidative stress. We describe steps for isolating mitochondria, preparing the dual-tube system, and encapsulation with lipid coat. We then detail procedures for performing assays and analyses. For complete details on the use and execution of this protocol, please refer to Nakano et al. 1 • Preparation steps for the mixture of mitochondria and lipids • Instructions for dual-tube-mediated coating • Handling guidance for lipid-coated mitochondria • Procedures to enhance intracellular ATP production under oxidative stress Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Mitochondria transplantation therapy is emerging as a novel therapeutic approach to promote neuroprotection in central nervous system (CNS) disorders. Here, we present a protocol for mitochondrial surface modification that enhances the restoration of intracellular ATP levels under conditions of oxidative stress. We describe steps for isolating mitochondria, preparing the dual-tube system, and encapsulation with lipid coat. We then detail procedures for performing assays and analyses.","journal":"STAR Protocols","year":2025,"id":537022,"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":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9576,"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":254198,"name":"David Back","orcid":"0000-0002-7381-4799","position":1,"is_corresponding":false},{"id":1052012,"name":"Ester Licastro","orcid":"0000-0001-6917-4605","position":2,"is_corresponding":false},{"id":1399733,"name":"Shin Ishikane","orcid":null,"position":3,"is_corresponding":false},{"id":868441,"name":"Takafumi Nakano","orcid":"0000-0001-5667-1979","position":4,"is_corresponding":false},{"id":279632,"name":"Kazuhide Hayakawa","orcid":"0000-0002-1229-2413","position":5,"is_corresponding":false},{"id":383535,"name":"Gen Hamanaka","orcid":"0000-0003-0957-2817","position":0,"is_corresponding":true}],"reference_count":4,"raw_metadata":null,"created_at":"2026-07-19T02:52:09.056872Z","pmid":"41175371","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":[]}