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Variability in success rates in literature and significant ethical concerns with animal welfare necessitate further optimization of adriamycin (ADR)\u2013induced FSGS model developed on BALB\/c mice.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>High\u2010performance liquid chromatography (HPLC) was used to assess ADR stability in water and upon light exposure. To identify the optimal ADR level, single intravenous ADR injections with dosing levels from 10 to 17\u2009mg\/kg body weight were administered to BALB\/c mice to induce FSGS\u2010like pathology. Body weight and proteinuria of FSGS mice were monitored and analyzed for FSGS model\u2013associated morbidity. Animals were euthanized for hematological and kidney histological assessments 8\u2009weeks post induction. To identify the suitable experiment time frame of the ADR\u2010induced FSGS mouse model, a longitudinal study was performed, with an 11\u2010week continuous monitoring of the symptoms.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>ADR was found to be unstable in aqueous media and light sensitive. A dosing level of 10.5\u2009mg\/kg of ADR was optimal for consistent FSGS mouse model induction on BALB\/c strain, characterized by minimal mortality and sustained FSGS\u2010like symptoms. Findings from the longitudinal study suggest that 6\u2009weeks post ADR induction may represent the peak of FSGS pathology severity in this mouse model. This time frame may be used for FSGS drug development projects.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Based on the outcome from this study, we identified the optimal ADR dosing level and model testing duration. A standard operating procedure (SOP) for the ADR\u2010induced FSGS mouse model was established to facilitate FSGS basic research and drug development.<\/jats:p><\/jats:sec>","DOI":"10.1002\/ame2.12564","type":"journal-article","created":{"date-parts":[[2025,2,7]],"date-time":"2025-02-07T09:44:46Z","timestamp":1738921486000},"page":"854-863","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Refining the adriamycin\u2010induced focal segmental glomerulosclerosis mouse model to improve reproducibility and animal welfare"],"prefix":"10.1002","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7678-7992","authenticated-orcid":false,"given":"Haochen","family":"Jiang","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical Sciences, College of Pharmacy University of Nebraska Medical Center  Omaha Nebraska 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