Please use this identifier to cite or link to this item: https://www.um.edu.mt/library/oar/handle/123456789/34426
Title: Optimization of a flow cytometric method to confirm the differentiation of HL-60 cell lines by candidate chemicals as indicated by screening tests
Authors: Camilleri, Roberta
Keywords: Myeloid leukemia
Stem cells
Immunophenotyping
Flow cytometry
Issue Date: 2017
Citation: Camilleri, R. (2017). Optimization of a flow cytometric method to confirm the differentiation of HL-60 cell lines by candidate chemicals as indicated by screening tests (Master's dissertation).
Abstract: Submitted to the Faculty of Medicine and Surgery of the University of Malta in partial fulfilment for the degree of Master in Biomedical Science Acute Myeloid Leukemias (AMLs) are a group of heterogenous haematopoietic disorders. They are characterized by the accumulation of myeloid blasts in the blood and bone marrow resulting in haematopoietic insufficiency with or without leucocytosis. This project is a follow up of collaborative research as a part of a COST Consortium called STEM CHEM. The main aim of this collaboration is to work with chemicals and natural products which induce differentiation of the HL-60 leukaemic cell line. The main aim of this study is to identify the lineage and stage of differentiation of treated HL-60 leukemic cells according to shifts in their respective immunophenotype as the cell differentiates using Flow Cytometry. A panel of antibodies was designed to identify the lineage and maturing pattern of treated HL-60 cells. The panel consists of eight different antibodies: CD45 (gating marker); CD64 and CD 14 (monoytic markers); CD34 (blast marker); CD15 and MPO (granulocytic markers); and CD13 and CD11b (maturing markers). The staining method and the panel of antibodies were optimized. A set of candidate chemicals were tested to identify any shifts in the immunophenotype of the HL-60 cells. The panel of antibodies shows clear shifts in the immunophenotype of cells and therefore the chemicals which induced differentiation (shift in the immunophenotype) were clearly identified. The chemicals tested in this study resulted as showing approximately a PV of 70% between the standard screening tests and the gold standard technique flow cytometry. This study concluded that using screening tests to select the best candidate chemicals for differentiation is a feasible option, however, flow cytometry is important to carry out in order to confirm any shifts in the immunophenotypic pattern of cells and thus determine differentiation.
Description: M.SC.BIOMED.SCI.
URI: https://www.um.edu.mt/library/oar//handle/123456789/34426
Appears in Collections:Dissertations - FacM&S - 2017
Dissertations - FacM&SPat - 2017



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