The conjunctiva is a complex tissue that covers the eye beginning at the corneal limbus and extending over the inner surfaces of the eyelids. Due to its important functions in maintaining the health of the ocular surface, adequate in vitro models of conjunctival structure and function are essential to understand its roll in different pathologies. Because there is scarcity of human conjunctival tissue that can be used in research, cell lines are often the only option for initial studies. An immortalized human conjunctival epithelial cell (IM-HConEpiC) line is now commercially available; however, it is not very well characterized. In this study, we have developed a new protocol to culture these cells without the use of collagen-coated culture surfaces, but with a defined cell culture medium. We characterized IM-HConEpiCs cultured under these conditions and corroborated that the cells maintained a conjunctival epithelial phenotype, including acidic and neutral mucins, junctional proteins E-cadherin and zonula occludens 1, and expression of CK8 and CK19, among others. In addition, we analyzed the response to oxidative stress and inflammatory stimuli and found that IM-HConEpiCs respond as expected for conjunctival epithelial tissue. For instance, cells exposed to oxidative stress increased the production of reactive oxygen species, and that increase was blocked in the presence of an antioxidant agent. In addition, after stimulation with TNF-α, IM-HConEpiCs significantly increased the production of IL-1β, IL-6, IL-8, and IP-10. Therefore, with this study we conclude that IM-HConEpiCs can be a useful tool in functional studies to determine the response of the conjunctiva to pathological conditions and/or to test new therapeutic strategies.
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