Methods Development for the Isolation and Culture of Primary Corneal Endothelial Cells

The vesicating agent sulfur mustard (SM) continues to represent a worldwide threat because of the compound's ease of production, minimal cost of manufacturing, widespread stockpiling and the lack of definitive medical treatment. The vast majority of SM exposure cases involve corneal injury. At low-exposure doses, ocular irritation and inflammation will resolve within weeks and generally will not recur. However, higher exposure levels may result in a second phase of pathology that can arise decades after initial symptoms have resolved. These patients experience repeated cycles of severe edema, corneal erosion, pathological angiogenesis and corneal scarring that can eventually result in blindness. The medical complications of late-onset ocular SM injury are collectively referred to as mustard gas keratopathy (MGK). Prevailing evidence suggests that late onset MGK may result from a deficit in corneal endothelial cells (CECs). If so, cellular therapy intended to replenish CECs lost to injury and age may represent an effective approach in the treatment or prevention of MGK onset. Here we discuss a systematic evaluation of in vitro CEC propagation methods. These investigations define optimized protocols for CEC culture expansion to pharmaceutically relevant cell numbers, enabling cell replacement therapy as a potential option in the treatment of MGK.