Hypoxia-Induced Oxidative Stress Assay

HLE-B3 cells purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) were grown as a monolayer in DMEM supplemented with 20% heat-inactivated fetal bovine serum (FBS) at 37 °C in a humidified atmosphere of 5% CO₂ and 21% O₂. Twenty-four h before the day of the experiment, cells were switched to hypoxic conditions (1% O₂ to mock physiological environment [15 (link)]). At 85–90% confluence, the cells were treated with the indicated concentration of H₂O₂ for 24 h.

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Wang S., Guo C., Yu M., Ning X., Yan B., Zhao J., Yang A, & Yan H. (2018). Identification of H2O2 induced oxidative stress associated microRNAs in HLE-B3 cells and their clinical relevance to the progression of age-related nuclear cataract. BMC Ophthalmology, 18, 93.

Publication 2018

HLE-B3 cells

Atmosphere Cells Fetal bovine serum H2o2 Hypoxic Physiological

Corresponding Organization :

Other organizations : Air Force Medical University, Xijing Hospital, First Affiliated Hospital of Chongqing Medical University, Chongqing Medical University

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Variable analysis

independent variables

Concentration of H2O2

dependent variables

Response to H2O2 treatment

control variables

Cell culture conditions (DMEM supplemented with 20% heat-inactivated FBS, 37 °C, 5% CO2, 21% O2)
Cell confluency (85-90%)
Duration of H2O2 treatment (24 h)

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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