Modelling the gut barrier: going with the flow

Gut health is closely lined to gut barrier integrity and when the gut becomes “leaky” this allows the host to be exposed to bacteria and metabolites which can cause infection and inflammation. Gut cells, colonocytes, form a barrier between the host and gut microbes, and are thus central to maintaining gut integrity and health. This barrier is very important as it controls what the host and its organs are
exposed to, so molecules which can cause inflammation or bacteria which can cause sepsis are not able to move from the lumen of the gut into the host.

However, when modelling this barrier in the laboratory we use tissue culture systems which do not replicate what happens in real-life. Current systems do not include “flow” in their models and wherever you go in the human body cells are exposed to shear forces, e.g. in the gut or in veins and arteries. We have looked at using a new flow system developed by our PhD partner Kirkstall Ltd, which allows flow to be introduced as a feature of the tissue culture. Using this system we have shown that colonocytes, when grown in the presence or absence of flow, show significant differences in their physiology, e.g. they are more viable and produce more mucin with flow, but do not respond as strongly to an proinflammatory stimulus.

This studentship will be split into two work packages (WP) and will aim to further characterise the impact of flow on growing colonocytes and whether the flow system replicates real life in the gut.

WP1 will characterise different colonocytes cell lines under flow and non-flow conditions, using microscopy (TEM, Confocal), gene expression, cell health parameters (Muse assays) and other tests of cell physiology and structure.

WP2, will test these cells with bacterial isolates and pro and anti-inflammatory challenges to assess how they compare to biopsy samples.

The student will learn a wide range of tissue culture, microscopy, histology, gene-expression and molecular biology to understand how flow affects gut integrity. They will also work closely with Kirkstall to understand the flow dynamics and characteristics of the Quasi Vivo® system

Specific research questions:-

1: How does flow affect cell physiology?
2: How does flow affect cell anatomy?
3: Does flow change the cellular responses to stimuli?
4: Do flow systems show more similarity with real-life colonocytes?
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