{"doi":"10.1016/j.ymeth.2025.03.019","title":"Optimized toolkit for the manipulation of immortalized axolotl fibroblasts","abstract":"The axolotl salamander model has broad utility for regeneration studies, but this model is limited by a lack of efficient cell-culture-based tools. The Axolotl Limb-1 (AL-1) fibroblast line, the only available immortalized axolotl cell line, was first published over 20 years ago, but many established molecular biology techniques, such as lipofectamine transfection, CRISPR-Cas9 mutagenesis, and antibiotic selection, work poorly or remain untested in AL-1 cells. Innovating technologies to manipulate AL-1 cells in culture and study their behavior following transplantation into the axolotl will complement in-vivo studies, decrease the number of animals used, and enable the faster, more streamlined investigation of regenerative biology questions. Here, we establish transfection, mutagenesis, antibiotic selection, and in-vivo transplantation techniques in axolotl AL-1 cells. These techniques will enable efficient culture with AL-1 cells and guide future tool development for the culture and manipulation of other salamander cell lines.","journal":"Methods","year":2025,"id":563765,"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":0.9435,"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":1394289,"name":"Glory Kalu","orcid":null,"position":1,"is_corresponding":false},{"id":1466970,"name":"S. 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Luong","orcid":null,"position":12,"is_corresponding":false},{"id":1393717,"name":"Aaron M. Savage","orcid":"0000-0002-2636-6145","position":13,"is_corresponding":false},{"id":492096,"name":"Sangwon Min","orcid":"0000-0001-9744-5847","position":14,"is_corresponding":false},{"id":1466976,"name":"Alparslan Karabacak","orcid":null,"position":15,"is_corresponding":false},{"id":1393722,"name":"Sebastian Böhm","orcid":"0009-0006-4599-6309","position":16,"is_corresponding":false},{"id":1279128,"name":"Ryan T. 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