Characterization of epithelial primary culture from human conjunctiva
Authors
L. Rivasa, A. Blázquez, F.J. Mu˜noz-Negrete, S. López, G. Rebolleda,F. Domínguez, A. Pérez-Esteban
Institution
Hospital Ramon y Cajal
Country
Spain
Year
2014
Journal
ARCHIVOS DE LA SOCIEDAD ESPAÑOLA DE OFTALMOLOGÍA
Abstract
Objective: To evaluate primary cultures from human conjunctiva supplemented with fetalbovine serum, autologous serum, and platelet-rich autologous serum, over human amnioticmembrane and lens anterior capsules.Methods: One-hundred and forty-eight human conjunctiva explants were cultured inCnT50®supplemented with 1, 2.5, 5 and 10% fetal bovine serum, autologous serum andplatelet-rich autologous serum. Conjunctival samples were incubated at 37◦C, 5% CO2and 95% HR, for 3 weeks.Results: The typical phenotype corresponding to conjunctival epithelial cells was presentin all primary cultures. Conjunctival cultures had MUC5AC-positive secretory cells, K19-positive conjunctival cells, and MUC4-positive non-secretory conjunctival cells, but werenot corneal phenotype (cytokeratin K3-negative) and fibroblasts (CD90-negative).Conclusions: Conjunctiva epithelial progenitor cells were preserved in all cultures; thus, a cellculture in CnT50®supplemented with 1–5% autologous serum over human amniotic mem-brane can provide better information of epithelial cell differentiation for the conjunctivalsurface reconstruction.