Establishment of optimal culture media in corneal epithelial wound healing models

Abstract

BACKGROUND: Wound healing needs to occur after injury to prevent vision loss. Models of wound healing need to be optimized to assure treatments for corneal wounds can be developed in vitro prior to investigating with in vivo studies. OBJECTIVE: The purpose of this study was to establish the optimum media to use as a control solution in wound healing models. METHODS: Immortalized human corneal epithelial cells were cultured in different growth media using a scratch and exclusion zone model. The effect of normoxic and hypoxic conditions on tight junctional integrity and metabolic activity of cells grown in different growth medium were also investigated. RESULTS: Wound healing with DMEMF12 media was significantly faster than both Keratinocyte serum-free media (p <  0.05) and EpiLife (p <  0.05) after 10 hours recovery under normoxic or hypoxic conditions using the scratch model and 9 days after wounding using the exclusion zone technique (p <  0.05). Using the culture media DMEMF12, cells stained for abundant ZO-1, Cx43 and had a high metabolic activity indicating significant epithelial barrier formation, gap junction formation and high cell viability. CONCLUSIONS: DMEMF12 led to superior wound healing under hypoxic and normoxic conditions and in two different wound healing models.