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Title: Development of novel endothelial cell culture systems and their applications in studying angiogenesis modulators from medicinal plants
Author: Cho, C. C.
Awarding Body: University of Cambridge
Current Institution: University of Cambridge
Date of Award: 2011
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The impact of the different culture media on human umbilical vein endothelial cells (HUVECs) and their responses to external treatments were examined. A new modified medium, MM, was developed by modifying a widely used endothelial cell growth medium – EGM-2. MM allows detection of the stimulatory effects of VEGF and other pro-angiogenic substances in the MTS cell proliferation assay and HUVEC-fibroblast tube formation assay and remains suitable for testing anti-angiogenic substances. Culture medium has a significant impact on HUVECs’ responsiveness to angiogenesis modulators and their consequent signalling studies. Therefore, simultaneous testing of potential modulators in a duo-medium system (i.e. MM and EGM-2) can minimise false negatives and experimental artifacts in preliminary screen. A volatile oil and two compounds derived from Angelica sinensis produced non-overlapping concentration-dependent responses on HUVEC proliferation and tube-formation assays. Furthermore, Angelica polysaccharides also showed immuno-modulatory property and induced proliferation of human peripheral blood mononuclear cells (PBMCs) in vitro whilst the PBMC-conditioned media showed potent pro-endothelial and pro-angiogenic effects in vitro. Through the incorporation of an immune component in the angiogenesis assays in vitro, a more comprehensive examination of natural products is possible. To explore the feasibility of using metabolomics to study angiogenesis, I used 1H-NMR to monitor the metabolic profile of endothelial injury and regeneration in an in-vitro model of wound healing. Preliminary data showed the most significant changes occurred between 0 – 6h post-injury, with increased levels of lactate, glutamine and choline accompanied by decreased levels of glutamate, and ATP. However, further refinements of this approach are necessary to establish a comprehensive view of the cellular activities associated with distinct phases of angiogenesis.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available