Application of a human primary epithelial-fibroblast co-culture system to study fibrosis

Richardson, Jaineeta (2017) Application of a human primary epithelial-fibroblast co-culture system to study fibrosis. PhD thesis, University of Nottingham.

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Idiopathic Pulmonary Fibrosis (IPF) is a chronic, progressive lung disease which manifests over several years with an unknown aetiology. The current paradigm suggests that reoccurring injury to the lungs results in damage to the alveolar epithelial cells, progressing to the destruction of the alveolar-capillary basement membrane. Although little is known about how the alveolar epithelium is injured, it is thought that repeated epithelial injury releases a wide variety of soluble factors which can initiate a series of events such as migration, differentiation and activation of mesenchymal cells i.e. fibroblasts into myofibroblasts. Myofibroblasts are proliferative, protected from apoptosis and produce extra-cellular matrix (ECM) in IPF. In turn it is likely these mesenchymal cells modify epithelial behaviour, producing a pro-fibrotic environment. Dysregulation of the normal processes in the lung results in abnormal communication between epithelial and mesenchymal cells which prevents resolution of the wound-repair response leading to tissue fibrosis and eventual organ failure.

In this project a 3D co-culture model was designed to investigate the interactions between epithelial cells and fibroblasts. This type of method enabled a non-contact approach between epithelial cells and fibroblasts in collagen mimicking the in vivo micro-environment. In the model, repetitive epithelial injury resulted in increased fibroblast proliferation, ECM synthesis and decreased apoptosis, similar to the biology of IPF. Affymetrix analysis revealed up regulation of fibroblast genes involved in cell cycle, proliferation, differentiation and in regulation of transcription in response to epithelial injury. Potential mediators of epithelial-fibroblast cross-talk were identified and revealed endothelin-1 to be involved in increased fibroblast proliferation and ECM gene expression. Treatment with bosentan against endothelin receptors resulted in the inhibition, in which there was reduced proliferation and matrix synthesis in the co-culture fibroblasts. Using the 3D co-culture model revealed that injury to epithelial cells can influence the response to fibroblasts in vitro. This then can be used with further antagonists, to study various mediators and block different pathways. We suggest that this model can be reliably used as a tool to investigate the interactions between various cell types.

Item Type: Thesis (University of Nottingham only) (PhD)
Supervisors: Johnson, S.R.
Clements, Debbie
Keywords: Pulmonary fibrosis; Epithelial cells; Fibroblasts; Cell interactions
Subjects: W Medicine and related subjects (NLM Classification) > WF Respiratory system
Faculties/Schools: UK Campuses > Faculty of Medicine and Health Sciences > School of Medicine
Item ID: 46427
Depositing User: Richardson, Jaineeta
Date Deposited: 15 Dec 2017 04:40
Last Modified: 03 May 2018 23:15

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