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Title: Haematopoietic differentiation of murine embryonic stem cells
Author: Gordon-Keylock, Sabrina Anne Megan
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2009
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This thesis aimed to determine which subregions of the E10.5 aorta-gonad-mesonephros (AGM) were responsible for the haematopoietic enhancing effects that primary AGM regions had on differentiating ES cells. To this end, a novel co-culture system has been established to test the enhancing effects of a panel of clonal stromal cell lines derived from different subregions of the midgestational AGM. Three stromal cell lines derived from the dorsal aorta and surrounding mesenchyme (AM) subregion of the AGM were able to significantly enhance the frequency of ES cell derived multipotent haematopoietic progenitors. Two stromal cell lines derived from the urogenital ridges (UG) of the AGM did not enhance haematopoietic differentiation of ES cells. The haematopoietic enhancing effects were not retained by extracellular matrices isolated from the AM stromal cell layers and the effects were dependent on direct ES cell-stromal cell contact. Co-culture of an ES cell line carrying a Brachyury-eGFP reporter gene demonstrated that the stromal lines mediated their effects post-Brachyury (mesoderm) induction in the ES cells. In addition, co-culture of sorted ES cell populations confirmed that Brachyury+, but not Brachyury-, cells gave rise to haematopoietic progenitors in AM co-culture, supporting the notion that ES cell differentiation recapitulated the in vivo pattern of lineage specification. Transplantation of co-cultured ES cells into irradiated adult NOD/SCID mouse recipients led to low levels of engraftment in the spleen and bone marrow. Adult bone marrow cells achieved repopulation more readily in the NOD/SCID animal model when transplanted intra-splenically, compared to intravenous injection. This suggests that transplantation of ES-derived haematopoietic cells directly into the haematopoietic niche, by intra-splenic or intra-femoral injection, could facilitate repopulation.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available