Oxygen-Glucose Deprivation/Reoxygenation (OGD/R) for in vitro Cerebral I/R

OGD/R has been recognized as an in vitro model for simulating I/R injury (Chen et al., 2020). PC12 cells have been widely used in neurophysiological and pathological research (Koubi et al., 2005). To mimic cerebral I/R in vitro, PC12 cells were subjected to OGD/R according to prior demonstrations (Ren et al., 2019). Briefly, cells were washed three times with phosphate-buffered saline (PBS; Gibco, Gaithersburg, MD, USA) and incubated for 12 h in glucose-free Dulbecco’s modified Eagle’s medium (DMEM; Gibco, Gaithersburg, MD, USA) medium under hypoxic conditions (1% O₂, 94% N₂, and 5% CO₂). The cells were then incubated under normoxic conditions (95% air and 5% CO₂) in RPMI-1640 medium for 1 h for reoxygenation. The cells were cocultured with BMSC-Exos (10 μg/ml), chloroquine (Cq; Sigma, Georgetown, SC, USA; 5 μM) or compound C (Sigma, Georgetown, SC, USA; 5 μM) during OGD/R.