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Title: Development of an ex vivo three dimensional (3-D) model of acute myeloid leukaemia (AML)
Author: Mortera Blanco, Teresa
Awarding Body: Imperial College London
Current Institution: Imperial College London
Date of Award: 2009
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Acute Myeloid Leukaemia (AML) is a cancer of hematopoietic stem cells that develops in the three-dimensional (3-D) niches provided by the bone marrow microenvironment in vivo. The study of AML has been hampered by the lack of appropriate ex vivo models, which can mimic this microenvironment. It was hypothesised that the fabrication of scaffolds for the biomimetic growth of leukemic cells ex vivo could facilitate the study of the disease in its native 3-D niche. The growth of different leukemic cell lines was first evaluated, namely K- 562, HL-60 and Kasumi-6 on highly porous scaffolds fabricated from biodegradable and non-biodegradable polymeric materials: poly (L-lactic-co-glycolic acid) (PLGA), polyurethane (PU), poly (methyl-methacrylate) (PMMA), poly (D, L-lactade) (PDLLA), poly (caprolactone) (PCL), and polystyrene (PS). These results were compared with two commercially available scaffolds from BD™ Biosciences. Overall, out of all the scaffolds, PLGA and PU displayed the best seeding efficiency and leukemic cellular growth, assessed by MTS assay, scanning electron microscopy and immunohistochemistry. In order to improve the ex vivo 3-D leukemic cell culture, PLGA and PU scaffolds were coated with bone marrow extracellular matrix (ECM) proteins, collagen (62.5 or 125 μg/ml) and fibronectin (25 or 50 μg/ml) and a combination of both proteins: collagen + fibronectin (62.5 + 25 μg/ml) respectively. Once the abnormal hematopoietic 3-D model was established, a new model to culture normal hematopoietic cord blood mononuclear cells was studied and compared. All 3 leukemic cell lines and cord blood cells grew better in PU scaffolds coated with collagen type I using the low concentration and sustained growth in the absence of exogenous cytokines. As a result, it was concluded that PU-collagen scaffold could provide a practical model with which to study the biology and treatment of primary AML in an ex vivo mimicry without the use of animal models.
Supervisor: Mantalaris, Sakis ; Panoskaltsis, Nicki Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral