2.2 Cell grouping and the hyperoxia model
This experiment included four groups. The control group featured primary
HUVECs in ECM medium which were cultured at 37℃ in a 5%
CO2 incubator. The Normoxia+Harmine group featured
primary HUVECs cultured in ECM medium containing harmine (at a final
concentration of 3μmol/L) under the same conditions as the control
group. In the Hyperoxia group, primary HUVECs were added to ECM medium;
then we applied the hyperoxia modeling method described in our previous
study 23. The cells were treated with a high-purity
gas mixture containing 950 ml/L O2 and 50 ml/L
CO2 at a rate of 3 L/min for 10 min. The oxygen
concentration was measured dynamically with a ML-IICB digital
intelligent oxygen system and maintained above 90%. The cells were
treated with high levels of oxygen once every 24 hours, and then
cultured under the same conditions as the control group. In the
Hyperoxia+Harmine group, the primary HUVECs were cultured in ECM medium
containing harmine at a final concentration of 3μmol/L under the same
conditions as the hyperoxia group. The next steps of the experiment were
carried out after the cells in the four groups had been cultured for 48
hours.