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Title: Embryonic stem cell differentiation : a novel approach to gene targeting in myeloid cells
Author: Smith, Aileen M.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2005
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This thesis demonstrates that macrophages can be generated from the culture of ES cells. Although low in number these macrophages show similar morphology and surface phenotype to bone marrow derived macrophages. Importantly, these ES macrophages readily phagocytosed latex beads and apoptotic cells. In additions, ES cells produced cells with a dendritic cell phenotype that were capable of apoptotic cell phagocytosis and maturation. Generation of these myeloid cells from ES cells was strongly dependent on serum and the parent ES cell line. Dendritic cells could be generated from integrin αv disrupted ES cells. These DCs retained the ability to phagocytose apoptotic cells suggesting that integrin αv is not essential for this process. This study also investigates the properties of Cre fused to two transduction proteins. Although Cre retained its recombinase activity as a fusion protein these were unable to translocate into cells. However Cre could be delivered to both primary macrophages and cultured epithelial cells using a replication deficient adenovirus allowing deletion of the targeted genes. Intriguingly, the apoptotic cell phagocytosis was unaffected by lack of integrin αv but could no longer be inhibited by the integrin antagonistic peptide RGD. In conclusion, the capacity of ES cells to differentiate to myeloid cells combined with the ability to deliver Cre to silently targeted myeloid cells provide powerful systems for studying the role of specific genes in phagocytosis. Use of these approaches with integrin αv demonstrates that this gene is not essential for apoptotic cell phagocytosis. However the ability of specific antagonists to inhibit phagocytosis show that integrin α­v is intimately involved in this process in normal cells.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available