3D co-culture angiogenesis assay
As a second-year veterinary student, receiving the AVTRW Diamond Jubilee award proved to be an invaluable step towards gaining essential research experience. The support provided a great opportunity to undertake a project that extended my curricular learning in a research setting and reinforced my interest in veterinary research.
The 2D tube formation and scratch wound assays, previously established to measure the angiogenic potential of endothelial cells(EC) in the horse, provide useful correlates for EC activity. Both assays, however, remain distant from the in vivo situation.
My project, ‘Development of a 3D co-culture angiogenesis assay using equine cells’, aimed to optimise assay protocols for use with equine ECs to facilitate mechanistic investigation of vascular equine EC function in a system that better recapitulates the physiological environment. The protocols for co-culture were optimised using ECs and fibroblasts, and the effects of pro-angiogenic growth factors, FGF-2, VEGF, and IGF-1, on vessel formation in the assay were assessed.
Analysis revealed that a seeding density of 1.6 × 104 ECs per well-established initial baseline vessel formation. Biologically relevant responses to putative pro-angiogenic factors validated the utility of the assay to model molecular control of equine angiogenesis. Our findings provided direction for future optimisation and demonstrated the potential of the co-culture assay as a robust model for mechanistic studies of angiogenesis in a system that better resembles tissue organisation.
Presenting this project at the AVTRW conference as a wonderful experience – made especially memorable by the encouraging atmosphere. The breadth of topics discussed also provided good insights into current directions within the various arms of the veterinary profession.