Method Profile
Compare five functional stress/injury endpoints across compound-specific concentration ranges in differentiated human proximal-tubule monolayers.
Mechanistic screening and prioritization within a focused platinum-drug kidney-injury case study.
One conditionally immortalized human proximal-tubule epithelial cell line; static differentiated monolayers.
Mechanistic exposures: cisplatin 5/10/20 μM, oxaliplatin 15/30/60 μM and carboplatin 60/120/240 μM, each with untreated controls; 24/48/72-hour injury measurements and a separate 2–48-hour ROS time course. These selected ranges are distinct from PrestoBlue viability range-finding and the 72-hour IC50 estimates.
Cells and compounds
ciPTECs supplied by Cell4Pharma; cisplatin, oxaliplatin and carboplatin; at least three independent biological repeats with three technical replicates per condition.
Endpoint panel
PrestoBlue metabolic activity, JC-10 mitochondrial potential, fluorescent ROS probe, LDH leakage and extracellular NAG activity; matched controls and endpoint-appropriate positive controls where reported.
Compare endpoint sensitivity, potency and time pattern within each drug’s selected range. Injury-event ordering is descriptive; no quantitative key-event relationship model was fitted.
Profile three related platinum agents with different cytotoxic potencies across linked stress/injury assays.
Single cell line; three compounds; at least three biological repeats and technical triplicates per condition.
Time-matched untreated/vehicle controls, compound-specific viability ranges and assay positive controls; no separate lysosome-specific positive control.
72-hour viability IC50: cisplatin 10.5 μM, oxaliplatin 45.5 μM, carboplatin 97.0 μM. Mitochondrial effects were generally most sensitive; reactive oxygen species (ROS) were robust for cisplatin, modest/transient for oxaliplatin and minimal/inconsistent for carboplatin. NAG largely tracked late membrane injury.
Human proximal-tubule cells showed distinct responses to cisplatin, oxaliplatin and carboplatin across five assay endpoints. Mitochondrial depolarization was generally more sensitive than overt injury, while oxidative-stress responses varied and lysosomal-enzyme release largely tracked later membrane damage.
