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Marie A. Shatos, Jose D. Rios, Vanja Tepavcevic, Harumi Kano, Robin Hodges, Darlene A. Dartt; Isolation, Characterization, and Propagation of Rat Conjunctival Goblet Cells In Vitro. Invest. Ophthalmol. Vis. Sci. 2001;42(7):1455-1464.
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purpose. To isolate, culture, and characterize goblet cells from the conjunctiva
methods. Conjunctival tissue was surgically removed from Sprague-Dawley rats.
Goblet cells were then isolated from the nictitating membrane and
fornix using explant cultures. Cells derived from the explants were
grown and propagated in RPMI medium supplemented with 10% fetal bovine
serum. They were characterized using an enzyme-linked lectin assay
(ELLA) with the lectin Ulex europaeus agglutinin-1
(UEA-1), Western blot analysis, PCR, light and electron microscopy,
specialized histochemistry and indirect immunofluorescence microscopy.
results. Goblet cells were successfully isolated from conjunctival explants by
scraping nongoblet cells from the culture vessel. To date, cultures
have been passaged a minimum of three times without the loss of their
specific cellular markers. Cells identified as goblet cells fulfilled
the following criteria: positive staining for alcian blue/periodic acid
Schiff reagent, cytokeratin (CK)-7, the lectins UEA-I and Helix
pomatia agglutinin (HPA), MUC5AC, and M3 muscarinic
receptor; detection of MUC5AC mRNA using RT-PCR; and negative staining
for CK-4, M1 muscarinic receptor, and Banderia
simplicifolia lectin. The authors also measured, using the
ELLA, substantial amounts of UEA-I–detectable high-molecular-weight
glycoproteins and MUC5AC released into the medium.
conclusions. Cultured goblet cells retain many characteristics of goblet cells in
vivo and thus may serve as a useful tool in delineating the
pathobiology of the ocular surface.
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