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M.T. Yen, L. Luo, D.–Q. Li, S.C. Pflugfelder; Phenotypic Characterization of Cultured Human Conjunctival Epithelial Cells Expanded Ex Vivo on Amniotic Membrane . Invest. Ophthalmol. Vis. Sci. 2004;45(13):1485.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To characterize the phenotype of cultured human conjunctival epithelial cells expanded ex vivo on human amniotic membrane. Methods: The phenotype of human conjunctival explants obtained from patients undergoing lower eyelid surgery was evaluated using hematoxylin–eosin, PAS, and immunostaining with antibodies against keratins (K3, K4, K7, K19), mucins (MUC–4, MUC–5AC), p63, and integrins α9 and ß1. Human conjunctival explants were then cultured on human amniotic membrane to subconfluence. Some cultures were air–lifted for an addition 7 days to promote stratification. The phenotype of the epithelialized amniotic membrane was then evlauated with histological staining and immunostaining. Results: Immunostaining of human conjunctival explants were K4, K7, and K19 positive but K3 negative. The basal layer of the epithelia stained positively to p63, integrin α9, and integrin ß1. The superficial layers were MUC–4 and MUC–5AC positive. Goblet cells were identified with PAS staining. The cultured conjunctival epithelia on amniotic membrane were noted to be stratified to 3–5 layers with uniformly small cells. They showed a similar phenotype to the conjunctival explants with K4, K7, and K19 positive but K3 negative immunostaining. Cultured epithelia expanded on amniotic membrane exhibit non–goblet epithelial cell differentiation. Conclusions: Human conjunctival epithelia expanded on amniotic membrane resembles the in vivo phenotype of human conjunctiva and may have therapeutic potential for conjunctival surface reconstruction.
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