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.

In vitro study of the cytotoxicity and antiproliferative effects of surfactants produced by Sphingobacterium detergens

“…Previous studies have found fibroblasts (embryonic origin) to be more sensitive than keratinocytes (epidermal origin). Benavides et al (2004a), Clothier et al (1999, Ree et al (1981) and Boukamp et al (1988) describe a higher resistance of keratinocytes, which is associated with their anatomical position, as they are constantly exposed to external aggression.…”

Ultrastructural Studies on the Effect of Photoactivated 8‐methoxy Psoralen

“…the cells were subcultured in Fdcon plastic tissue culture flasks (75 cm2) containing 13 ml Dulbecco's modified Eagle's culture medium supplemented with 2% foetal calf serum, 12% horse serum and antibiotics. The present study was carried out simultaneously with that previously reported ( 7 ) and the same serum lots were used.…”

“…The cells were fixed as described previously ( 7 ) and photographed as a monolayer on Tyrode buffer. One thousand cells from cultures fixed 2 hours, 3 days and 6 days after PUVA exposure were scored for the number of nuclear elements per cell.…”

“…We therefore considered it important to conduct further studies and have made an attempt to define the ultrastructu-ral alterations taking place in the nuclei of cells after PUVA exposure. As a model we have used an established human epithelial cell line which we have ultrastructurally and otherwise characterized in a preceding paper (7).…”

Morphology and Frequency of Heterochromatic Nuclear Sheets in Normal Circulating Human Leucocytes and after PUVA Therapy

“…followed by centrifugation at 1146 g for 15 min. Apart from unimportant details the cells were further processed as previously described by us (Ree et al 1981a). In short the cells were postfixed in 1 % buffered osmium-tetroxide, dehydrated, embedded in Epon 812, cut with a diamond knife and post-stained with uranyl acetate and lead citrate.…”