PURPOSE. TO isolate, culture, and characterize goblet cells from the conjun
ctiva of rats.
METHODS. Conjunctival tissue was surgically removed from Sprague-Dawley rat
s. Goblet cells were then isolated from the nictitating membrane and fornix
using explant cultures. Cells derived from the explants were grown and pro
pagated in RPMI medium supplemented with 10% fetal bovine serum. They were
characterized using an enzyme-linked lectin assay (ELLA) with the lectin Ul
ex europaeus agglutinin-l (UEA-1), Western blot analysis, PCR, light and el
ectron microscopy, specialized histochemistry and indirect immunofluorescen
ce microscopy.
RESULTS. Goblet cells were successfully isolated from conjunctival explants
by scraping nongoblet cells from the culture vessel. To date, cultures hav
e 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), M
UC5AC, and Mg muscarinic receptor; detection of MUC5AC mRNA using RT-PCR; a
nd negative staining for CK-4, M-1, muscarinic receptor, and Banderia simpl
icifolia lectin. The authors also measured, using the ELLA, substantial amo
unts of UEA-I- detectable high-molecular-weight glycoproteins and MUC5AC re
leased into the medium.
CONCLUSIONS. Cultured goblet cells retain many characteristics of goblet ce
lls in vivo and thus may serve as a useful tool in delineating the pathobio
logy of the ocular surface.