Establishing an effective 3D cell culture model for the study of infectious respiratory diseases

Infectious respiratory diseases pose serious threats to human and animal health due to their high mortality and morbidity rates. Lung structure and composition varies between species. However, large animal models, particularly ruminants, are especially useful models for studying host-pathogen interactions and the pathophysiology of both animal and human diseases(1).

Microfluidic organoid or ‘organs-on-a-chip’ platforms are a promising new group of micro-engineered models that recapitulate 3D tissue structure and physiology(2). Precision-cut lung slices are another widely used ex vivo model for visualizing the relationship between lung structure and function(3). A particular strength of 3D cell culture techniques is that they provide highly tractable experimental platforms. However, significant limitations of currently available systems include cell differentiation and their short lifespan in ex vivo culture.

Aims and objectives

The proposed research project aims to develop and optimize a functional, stable and reproducible 3D bovine cell culture model of the lung to study respiratory infectious diseases. Specifically, the student will develop and compare two different 3D culture systems for the future purpose of studying epithelial-immune cell interactions in response to infection: organoids employing a novel Organ-on-Chip technology and precision-cut lung slices using fresh tissue obtained from the Vet School’s post-mortem facility and on-site abattoir.

Methodology

• The alveolar epithelium will be seeded onto a thin biological and stretchable membrane produced from the umbilical cord of the neonatal ruminant as a supporting matrix. The membrane will be micropatterned via in-situ stem cell differentiation into custom-made micromolds and used to substitute the membrane currently employed in standard Organ-on-Chip devices.
• A cartilage graft and smooth muscle will be developed to support the growth of the respiratory epithelium of trachea and bronchi. Elastin will provide the support for the bronchioles.
• The respiratory epithelium and the alveolar epithelium will be grown on an air-liquid interface using ad hoc prepared maintenance medium that also contains monocytes produced from the umbilical cord, with the aim of populating the epithelium with resident macrophages.
• The culture system will be perfused with medium using innovative microfluidic techniques.
• The lung organoid produced will then be compared to precision cut lung tissue established in slice culture conditions using the same microfluidic system.

Both 3D culture systems will be monitored for 30 days using a range of lab techniques to evaluate cell maturation, differentiation, function and survival, including the development and/or maintenance of key innate immune system components. Finally cultures will be tested establishing an in vitro infection.

Apply for this project

This project will be based in Bristol Veterinary School.

Please contact bvs-pgrstudent-admin@bristol.ac.uk for further details on how to apply.

Apply now!