High resolution imaging and analysis of endothelial tubulogenesis and blood vessel formation

Salisbury, Victoria Alice (2017). High resolution imaging and analysis of endothelial tubulogenesis and blood vessel formation. University of Birmingham. Ph.D.

Preview

Salisbury17PhD.pdf
PDF - Accepted Version
Download (6MB)

Abstract

The process of angiogenesis in which new blood vessels form from pre-existing vessels, can be intensively studied through the use of \(in\) \(vitro\) and \(in\) \(vivo\) models. The \(in\) \(vitro\) co-culture tube formation assay is used to assess the ability of endothelial cells to develop into three dimensional tubular structures which mimics the growth of capillaries. Different fluorescent labelling techniques were developed and utilised alongside confocal microscopy to visualise endothelial tubulogenesis and investigate the mechanisms of lumenogenesis. Imaging the actin cytoskeletal organisation by expressing the lifeact peptide conjugated to fluorescent proteins revealed that Factin fibres outline lumens within endothelial tubules and enabled clear visualisation of filopodia formation.
Further studies presented in this thesis aimed to develop, test and evaluate computational tools for analysing endothelial sprouting from fluorescently labelled spheroids generated using the \(in\) \(vitro\) hanging drop spheroid assay and quantify blood vessel formation in the \(in\) \(vivo\) zebrafish model. The results confirmed that both analysis tools were able to rapidly quantify a wide range of angiogenic images and generated comparable results to frequently used manual methods. The developed computational analysis tools are user friendly and can be used to assess the effects of inhibitor compounds and silencing vascular related genes.

Type of Work:

Thesis (Doctorates > Ph.D.)

Award Type:

Doctorates > Ph.D.

Supervisor(s):