Egf phg0311

For the isolation and culture of hAECs, the amnion was mechanically peeled from the underlying chorion and washed three times with phosphate-buffered saline (E607008, PBS, Sangon Biotech, China) supplemented with 10% penicillin and streptomycin (15070063, Thermo Fisher Scientific, USA). The amnion was incubated with 0.25% trypsin-EDTA (25200072, Thermo Fisher Scientific, USA) for 15 min at 37°C, and the human amniotic epithelial cells were mechanically detached from the amnion. Dispersed hAECs were collected via centrifugation at 1500 rpm for 10 min, plated in 10-cm cell culture dishes in DMEM/F-12 medium (11330032, Thermo Fisher Scientific, USA) supplemented with 10% fetal bovine serum (FBS, ExCell Bio, China), 10 ng/mL human recombinant epidermal growth factor (EGF, PHG0311, Thermo Fisher Scientific, USA), 5 μM SB431542 (301836-41-9, Selleck, China) and 1% penicillin and streptomycin, and cultured in a cell incubator at 37°C. The hAEC culture medium was changed every 2 days.
Human corneal epithelial cells (hCECs) and human corneal stromal cells (hCSCs) were a kind gift of professor Qingjun Zhou (Shandong Eye Institute, China) and cultured in DMEM/F12 medium supplemented with 10% FBS in a cell incubator at 37°C.