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Description
Holoclone-, meroclone- and paraclone-like colonies were identified in in vitro cultures of normal epithelial cells. In the present study we tested whether cytokeratin 14 (ck14) and sonic hedgehog (shh) expression profiles are correlated with three buccal epithelial clone types. Buccal epithelial cells were isolated from porcine buccal tissues obtained from 9 donors. Cells were seeded into 12-well cell culture plates with glass coverslips in density of 2x104 cells/cm2. Three different clone types: holoclones, meroclones and paraclones were identified by two independent investigators based on their morphology. To characterize and compare the clones the immunofluorescent stains for ck14 and shh were performed. Fluorescence intensity of individual cells, cell surface and fluorescence intensity of background were measured to calculate relative cell fluorescence (RCF). The strongest expression of ck14 and shh was observed in the holoclones (201,4±78,52 and 9,23±3,26 respectively; p<0,05). The comparable ck14 expression was observed in the mero- and paraclones (p>0,01). Meroclones expressed significantly lower levels of shh (2,52±0,94) compared to paraclones (4,31±1,3; p<0,001). This study showed that three buccal epithelial clonal types with differing properties can be isolated from buccal tissues. Holoclones are the richest in shh (+) stem cells and this cell population should appear to be a promising alternative for obtaining urothelial cells from external urinary bladder sources.
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