In English

Wound healing in an in vitro bronchial epithelial cell model

Andreas Carlsson
Göteborg : Chalmers tekniska högskola, 2014. 98 s.
[Examensarbete på avancerad nivå]

The aim of this project was to investigate the potential use of differentiated primary human bronchial epithelial cells in air liquid interface as a model for wound healing in vitro. Wounds were simulated by mechanical scratching and whole cigarette smoke exposure. The outcome of the project is aimed to increase the knowledge of primary bronchial epithelial cells in the respiratory research, the ability to use the model for wound healing studies and the potential use of the model for compound testing. The primary read-outs used to evaluate the model were real-time quantitative polymerase chain reaction, immunocytochemistry, live imaging, image analysis and trans epithelial electrical resistance. From this project primary human bronchial cell cultures were concluded to have a high recovery potential to both scratch wounds and exposure of whole cigarette smoke. The wound healing results obtained from this project were connected to previous described theories of in vivo wound healing and the major cell populations were observed to recover over time. Cigarette smoke was observed to significant delay the wound healing of scratch wounded cell cultures, a feature used to investigate pharmaceutical compounds effect to migration in the cell model. Treatment of three different compounds to the model in this project did not show any effect to the cell migration. Live imaging of scratch wounded cell cultures revealed a migration within the whole cell culture during wound healing. Surprisingly, non-wounded fully differentiated cell cultures showed a constant cellular migration which, as to the writer's knowledge, has not been previous reported. Through this project a successful wound healing model was established using a combination of read-outs. A model for cigarette smoke exposure of air liquid interface cultures was performed and a window for compound treatment was discovered as a result of the impact of cigarette smoke on cellular migration. This project was performed in collaboration between Chalmers University of Technology and AstraZeneca R&D, at the section of Cell and Molecular Pharmacology within the department of Respiratory, In ammation and Autoimmunity at AstraZeneca R&D facility in Mölndal.

Nyckelord: Primary normal human bronchial epithelial cells, air liquid interface, wound healing, ICC, real-time qPCR, live imaging, confocal microscopy, cigarette smoke.



Publikationen registrerades 2014-06-05. Den ändrades senast 2015-01-19

CPL ID: 198934

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