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Epithelial tight junction function in a co-culture model of the human bronchial epithelium and endothelium

Epithelial tight junction function in a co-culture model of the human bronchial epithelium and endothelium
Epithelial tight junction function in a co-culture model of the human bronchial epithelium and endothelium

Epithelial cells line the conducting airways of the bronchial tract, with endothelial-lined capillaries in the submucosa, forming an integrated system and acting as a barrier to the environment.  When the epithelial surface is challenged, the epithelial and endothelial response is locally co-ordinated to allow infiltrating cells to cross the endothelium and target the site of challenge on the luminal surface.  This thesis reports the establishment of a co-culture model reflecting this co-ordinated system, to enable epithelial-endothelial cell interactions to be investigated.

16HBE 14o-, SV40-transformed bronchial epithelial cells were grown on the underside of insert membranes, and HUVEC (human umbilical vein endothelial cells) were grown on the upper side.  Immunofluorescent staining with HEA-125 (human epithelial antigen-125) provided an epithelial-surface marker and vWF (von Willebrand factor) an intracellular endothelial-specific marker.  Using these markers in confocal microscopy, showed that confluent epithelial and endothelial cell layers were growing on opposite sides of the membrane.  This was confirmed by scanning and transmission electron microscopy.  Transepithelial electrical resistance (TER) was used as a functional measurement reflecting tight junction formation.  The final model improves on existing models of the conducting airways, allowing the effects of cell-to-cell interaction on tight junction function to be investigated, better reflecting the in vivo situation in which epithelial and endothelial cells coexist.

The bilayer had an increased TER (1547Ω.cm2, S.E. ± 59) in comparison to the epithelial monolayer (792Ω.cm2, S.D. ± 84), and this increase only occurred when 16HBE 14o- cells were grown with endothelial cells.  Furthermore, this same increase was seen when HUVEC conditioned medium was added to the basolateral side of 16HBE 14o- cells, confirming that the increase in TER was occurring in the 16HBE 14o- cells.  These results revealed that functional differences exist between epithelial cells grown as a monolayer and epithelial cells grown as a bilayer with endothelial cells.

University of Southampton
Chowdhury, Ferdousi
0af499d4-17c5-40cf-9426-0d509ab82595
Chowdhury, Ferdousi
0af499d4-17c5-40cf-9426-0d509ab82595

Chowdhury, Ferdousi (2003) Epithelial tight junction function in a co-culture model of the human bronchial epithelium and endothelium. University of Southampton, Doctoral Thesis.

Record type: Thesis (Doctoral)

Abstract

Epithelial cells line the conducting airways of the bronchial tract, with endothelial-lined capillaries in the submucosa, forming an integrated system and acting as a barrier to the environment.  When the epithelial surface is challenged, the epithelial and endothelial response is locally co-ordinated to allow infiltrating cells to cross the endothelium and target the site of challenge on the luminal surface.  This thesis reports the establishment of a co-culture model reflecting this co-ordinated system, to enable epithelial-endothelial cell interactions to be investigated.

16HBE 14o-, SV40-transformed bronchial epithelial cells were grown on the underside of insert membranes, and HUVEC (human umbilical vein endothelial cells) were grown on the upper side.  Immunofluorescent staining with HEA-125 (human epithelial antigen-125) provided an epithelial-surface marker and vWF (von Willebrand factor) an intracellular endothelial-specific marker.  Using these markers in confocal microscopy, showed that confluent epithelial and endothelial cell layers were growing on opposite sides of the membrane.  This was confirmed by scanning and transmission electron microscopy.  Transepithelial electrical resistance (TER) was used as a functional measurement reflecting tight junction formation.  The final model improves on existing models of the conducting airways, allowing the effects of cell-to-cell interaction on tight junction function to be investigated, better reflecting the in vivo situation in which epithelial and endothelial cells coexist.

The bilayer had an increased TER (1547Ω.cm2, S.E. ± 59) in comparison to the epithelial monolayer (792Ω.cm2, S.D. ± 84), and this increase only occurred when 16HBE 14o- cells were grown with endothelial cells.  Furthermore, this same increase was seen when HUVEC conditioned medium was added to the basolateral side of 16HBE 14o- cells, confirming that the increase in TER was occurring in the 16HBE 14o- cells.  These results revealed that functional differences exist between epithelial cells grown as a monolayer and epithelial cells grown as a bilayer with endothelial cells.

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Published date: 2003

Identifiers

Local EPrints ID: 465350
URI: http://eprints.soton.ac.uk/id/eprint/465350
PURE UUID: c350e793-276f-4e98-b91b-7b17755884c0

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Date deposited: 05 Jul 2022 00:39
Last modified: 05 Jul 2022 04:59

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Contributors

Author: Ferdousi Chowdhury

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