May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Phenotypic Characterization of Cultured Human Conjunctival Epithelial Cells Expanded Ex Vivo on Amniotic Membrane
Author Affiliations & Notes
  • M.T. Yen
    Ophthalmology,
    Cullen Eye Institute, Baylor College of Medicine, Houston, TX
  • L. Luo
    Ophthalmology,
    Ocular Surface Center,
    Cullen Eye Institute, Baylor College of Medicine, Houston, TX
  • D.–Q. Li
    Ophthalmology,
    Ocular Surface Center,
    Cullen Eye Institute, Baylor College of Medicine, Houston, TX
  • S.C. Pflugfelder
    Ophthalmology,
    Ocular Surface Center,
    Cullen Eye Institute, Baylor College of Medicine, Houston, TX
  • Footnotes
    Commercial Relationships  M.T. Yen, None; L. Luo, None; D. Li, None; S.C. Pflugfelder, None.
  • Footnotes
    Support  NIH Grants EY11915 & EY014553, Research to Prevent Blindness, The Oshman Foundation, The Farish Fund
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 1485. doi:
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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)

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Abstract

Abstract: : 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.

Keywords: conjunctiva • immunohistochemistry • microscopy: light/fluorescence/immunohistochemistry 
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