The use of human embryonic stem cells to investigate human lens and cataract development

Abstract

The aim of this thesis was to improve the use of human pluripotent stem cells to provide a refined cell source, for lens and cataract research. Chapter 1 provides a general background to the impact of cataract and the complications associated with current treatment strategies, in order to highlight the need to develop novel anticataract treatments and the need for new lens models to aid future anti-cataract drug discovery. Chapter 2 was focused on optimising a published, though imperfect, differentiation system to generate a pure population of human embryonic stem cell derived lens epithelial cells that can be used in downstream anti-cataract drug screens. From the results of this chapter (i.e. the partial improvement in lens epithelial cell production) it was clear that greater knowledge of the regulation involved in the lens was needed to improve future differentiation methods. For this reason a simple bioinformatics tool was created and used in Chapter 3 to identify candidate new regulators of lens growth factor signalling pathways. The success obtained through Chapter 3 led to use of the bioinformatics tool in Chapter 4 to develop a method for purifying lens epithelial cells (via specific surface antigens) from mixed populations of lens fibre cells and non-lens cell types. Subsequent use of these purified human lens cells demonstrated a proof of principle for anti-cataract drug screening. Chapter 5 is a general discussion of the outcomes and benefits of this research and future initiatives for improvement. This thesis has been submitted in a composite format largely centred on the Investigative Ophthalmology and Visual Science (IOVS) manuscript format, however, section headers and numbering was added to aid reviewers. The Introduction of this thesis is an invited review manuscript, and the 3 included experimental chapters have been prepared for submission to IOVS (with delayed submission due to preparation and submission of the Australian Provisional Patent Application # 2014900269: A method for the purification of eye lens cells).

Date of Award	2014
Original language	English