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Title: Analysing the angiogenic potential of mesothelial cells in vitro and in vivo
Author: Ward, K. L.
ISNI:       0000 0004 6425 3795
Awarding Body: University of Liverpool
Current Institution: University of Liverpool
Date of Award: 2017
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In angiogenesis and arteriogenesis, vessel stabilisation through the recruitment of pericytes (PCs) and vascular smooth muscle cells (VSMCs) is crucial to enable vessels or collaterals to maintain integrity and support blood flow and pressure. Genetic lineage tracing studies in mice have shown the localisation of mesothelial cells (MCs) to forming vessels in the gut and heart. In our research we are keen to explore the angiogenic potential of murine-derived MCs with the aim to develop regenerative medicine therapies. This study used an in vitro angiogenesis assay, combined with live-cell and endpoint imaging, to explore the dynamics and interaction of MCs with human dermal microvascular endothelial cells (HDMECs). To track the cells and determine their localisation, they were labelled using a lentivirus-based approach. We have shown that MCs isolated from adult mice, have the capacity to migrate and align with endothelial tubes, with a higher affinity than PCs. The MCs had no impact on tube formation, as assessed by the area the tubes occupied and the number of branch points formed. We have also shown that aligning MCs expressed aSMA, a marker of PCs/VSMCs, suggesting that they differentiated towards a mural cell fate. To identify factors that could be responsible for inducing migration and subsequent alignment of MCs to the endothelial tubes, we performed several assays, including an angiogenesis array to explore up- or down-regulated proteins, and a Boyden chamber assay. We observed an increase in a number of angiogenesis-associated proteins using the angiogenesis array, including Angiopoietin-2, PIGF and interleukin-8. By contrast, the Boyden chamber assay revealed that MCs are attracted in a dose dependent manner by the two key angiogenic cues PDGF-BB and bFGF. We also explored the therapeutic effects of MCs in vivo using the pre-clinical mouse model of hind limb ischemia (HLI) induced by femoral artery ligation, and a sponge implant model. In both models, the MCs did not elicit a systemic effect. However, our analysis of the role of MCs in promoting and contributing to arteriogenesis and angiogenesis in the HLI model so far suggests that following femoral artery ligation, the MCs were unable to increase perfusion to the injured limb.
Supervisor: Wilm, B. W. ; Cross, M. ; See, V. Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral