DHA-promoted repair of human corneal epithelial cells in high-glucose environment

Abstract

Materials and methods MaterialsCorneal epithelial progenitor cells (TKE2) (ATCC); highglucose DMEM (Giboco); trypsin 0.25% (NanJing KeyGen Biotech); flow cytometry kit (NanJing KeyGen Biotech); Hoechst 33258 staining solution (Beyotime Institute of Biotechnology); PEDF, VEGF, Caspase-3, Caspase-9, and p-NF-κB (p65) antibodies (Abcam); PEDF, VEGF, Caspase-3, Caspase-9, and NF-κB (p65) primer sequence (NanJing KeyGen Biotech). Cell cultureAfter resuscitation, TKE2 cells were resuspended in complete medium (high-glucose DMEM+10% FBS) and placed in an incubator with 5% carbon dioxide at 37 °C. When the cells had grown to more than 80% adherently, subculture was conducted: The cells in the logarithmic growth phase were digested to prepare a single cell suspension, which was then inoculated in the culture plate at 1×104 / well and cultured in a cell incubator with 0.5% CO2 at 37 °C. After the cells were subcultured for one to two passages, they were subcultured to a total of 10 culture dishes of 35mm in diameter following the above method. The human corneal epithelial cells were stimulated with highglucose concentration (25 mmol/ L glucose in DMEM complete medium) to establish the Model group.