CHARACTERIZATION OF EPITHELIAL PRIMARY CULTURES FROM HUMAN CONJUNCTIVA

Primary cultures of human epithelial cells from normal conjunctiva wer e developed and characterized to determine whether they retained epith elial characteristics. Conjunctival explants were obtained from the up per fornix of healthy donors end cultured in supplemented DMEM/F-12 me dium for 5 days. The epithelial outgrowth was maintained for an additi onal IO days. Primary cultures were then processed for light microscop y, transmission and scanning electron microscopy (TEM, SEM), and immun ocytochemistry. They exhibited typical features of conjunctival epithe lium on light microscopy (polygonal morphology, intimate cohesion, pro duction of mucins), TEM (abundant desmosomes, keratin bundles, granule s, microvilli), SEM (polygonal shape, microvilli, intimate cohesion), and immunocytochemistry (positivity for the receptor of epidermal grow th factor, desmosomal proteins, and cytokeratins). In conclusion, prim ary cultures developed from normal human conjunctiva maintained the ep ithelial characteristics in vitro, Because the conjunctiva is a major component of the anterior ocular surface, we propose this in vitro sys tem as suitable for physiopathologic and toxicologic studies.