Characterization of a human epithelial cell line with special reference to its ultrastructure.

Abstract

A commercially available cell line (NCTC 2544) originating from presumably normal human skin was chosen as an in vitro model system for subsequent studies of the effects of different agents on human epithelial cells. The cell line, therefore, was ultrastructurally and otherwise characterized at intervals by techniques which allow standardized controls of the model. The cell line was classified as epithelial both by phase contrast microscopy and transmission electron microscopy. The cells were polygonal, fully developed desmosomes were demonstrated and no extracellular filamentous material was observed. Specific epidermal markers like keratohyalin granules or keratinosomes were not demonstrated. Comparison of the nuclear morphology of cells from 1, 3 and 6 days old cultures revealed that only minor changes took place. The same was true for cytoplasmic features. Thus this cell line has a well-defined ultrastructural morphology. Flow cytometry studies showed the cell line to be tetraploid, with no sub-populations with other ploidies. Surface antigens were typed by a microcytotoxicity assay using 12 different antisera. The electrophoretic patterns of the isoenzymes lactate dehydrogenase and glutathione reductase were human. The cells did not form colonies in soft agar. Though several parameters indicate that this cell line is not a HeLa cell contaminant, this possibility cannot, however, be totally excluded.