THE FATE OF GENETICALLY MARKED HUMAN ORAL KERATINOCYTES IN-VITRO

The fate of the progeny of human oral gingival keratinocytes was mapped in stratified epithelial tissues in vitro by following the expression of a marker gene in genetically related clones. Oral epithelial progenitor cells were genetically marked at high efficiency by transducing them with a retrovirus vector that carried the gene for a histochemically detectable product, Escherichia coli beta-galactosidase (beta-gal). These cells were then grown in submerged cultures and on collagen rafts at the air-liquid interface to demonstrate the distribution of genetically marked cells in a differentiating tissue in vitro. The dynamics of transduced cells showed that clonally related cells were arranged in discrete units of labelled cells and these clusters were defined as 'clonal proliferation units'. The size and configuration of these units were related to the proliferative potential and differentiating capacity of the cell that was initially transduced. This model demonstrates the relation between clonally related cells and tissue architecture for oral keratinocytes in vitro.