Hypoxia-Reoxygenation Model in HK-2 Cells

Immortalized human renal proximal tubule (HK-2) cells, with phenotypic and functional characteristics of proximal tubule cells, were obtained from the American Type Culture Collection (VA, USA). HK-2 cells were hatched in a culture system of Dulbecco's Modified Eagle Medium/F12 supplemented with 10% fetal bovine serum (Invitrogen, Carlsbad, USA), 100 U/mL penicillin G, 100 μg/mL streptomycin and 0.25 μg/mL amphotericin B (Invitrogen). Then, the cells were cultivated in a hypoxic environment (1% O₂, 94% N₂, and 5% CO₂) for 24 h and then in an aerobic environment (21% O₂, 74% N₂, and 5% CO₂) for 3 h to establish an in vitro H/R model [21 (link)].