A transformed human epithelial cell line that retains tight junctions post crisis

Abstract

The successful establishment of a postcrisis SV-40 T antigen transformed epithelial cell line, 1HAEo-, which retains tight junctions and vectorial ion transport, is described. Immunocytochemical analysis of 1HAEo- cells shows a defined pattern of cytokeratin staining and a characteristic pericellular localization of the adhesion molecule cellCAM 120/80, indicating the presence of junctional complexes. The presence of both tight junctions and desmosomes has been confirmed by electron microscopy. Cell monolayers have good transepithelial resistance measured in Ussing chambers. Cells increase chloride ion transport in response to addition of agents that raise either intracellular cAMP or calcium, measured either by 36Cl- efflux or whole-cell patch clamp. An increase in short-circuit current, in response to these agents, can be measured in Ussing chambers. The presence of a depolarization-induced outwardly rectifying 45 pS chloride channel has been demonstrated in single cell detached membrane patches. In addition, the cells have been found to express mucin mRNA. These cells therefore demonstrate that it is possible to select transformed cell clones with particular morphologic characteristics, i.e. the presence of tight junctions and cell polarity, which also retain useful epithelial cell-specific functions, including vectorial ion transport. They also provide a major resource for the study of the structure and function of human epithelia.

Comparative cytoprotective effects of carbocysteine and fluticasone propionate in cigarette smoke extract-stimulated bronchial epithelial cells

“…16-HBE, an immortalised normal bronchial epithelial cell line (Cozens et al 1992), RPMI 2650a, a nasal epithelial cell line (Pace et al 2012a), and a mucin-producing human NCI-H292 airway epithelial cell line (Nogawa et al 2009) were used in this study.…”

A comparison of linear and branched polyethylenimine (PEI) with DCChol/DOPE liposomes for gene delivery to epithelial cells in vitro and in vivo

“…We have compared the ability of three forms of PEI to deliver a reporter gene to a human bronchial epithelial cell line (16HBE) as a model for airway gene delivery under different culture conditions (subconfluent, confluent and polarized). 21,22 We have shown that all three forms of PEI gave significantly better reporter gene delivery compared to using liposomes. Linear 22 kDa PEI was significantly better than branched 25 or 50 kDa PEI.…”

“…The variation in response in the pT10CFTR2-treated animals meant that the average genistein response was not significantly different from the CF mice treated with the control plasmid pT10. If it is assumed, however, that the lack of a genistein response reflects a failure of gene delivery and these 'failures' are excluded, then the mean genistein response after L- 22 …”

“…Human bronchial epithelial cells 16HBE 22 (generously provided by Professor Dieter Gruenert, University of Vermont, VT, USA) were grown in minimal essential medium with Earle's salts (Invitrogen, Paisley, UK) supplemented with 10% foetal calf serum (FCS, TCS Biologicals, Buckingham, UK), 2 mM L-glutamine (Invitrogen), 100 U/ml penicillin and streptomycin (Invitrogen). Culture of polarized cells on Anopore membranest (Invitrogen) differed in that the FCS was replaced by 2% Ultroser G (Invitrogen).…”

Steroid hormone enhancement of gene delivery to a human airway epithelial cell line in vitro and mouse airways in vivo

“…We have used a human bronchial epithelial cell line (16HBE) as a model for airway gene delivery under several different culture conditions including sub-confluent, confluent and polarized. 15,16 We have shown significant enhancement of reporter gene delivery in each of these cell culture conditions using ␤-estradiol or methyl-prednisolone. The order in which steroid is combined in the complex is important.…”