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Identifier 000376420
Title Characterization of bone marrow mesenchymal stem cells from patients with myeloid disorders using chromosomal analysis and molecular genetic techniques
Alternative Title Χαρακτηρισμός των αρχέγονων μεσεγχυματικών κυττάρων του μυελού των οστών σε ασθενείς με μυελικές διαταραχές χρησιμοποιώντας χρωμοσωμική ανάλυση και τεχνικές μοριακής γενετικής
Author Batsali, Aristea
Author Μπάτσαλη, Αριστέα
Thesis advisor Παπαδάκη, Ελένη
Abstract In bone marrow (BM) there are at least two types of multipotent stem cells: hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs). Although in general these two cell types are distinct as regards their origin and differentiation potential, there are some indirect evidence supporting that HSCs and MSCs may have their origin from a common ancestor stem cell. The aim of this study is to investigate whether the abnormal clone in acute myeloid leukemia (AML) -, myelodysplastic syndromes (MDS) - or myeloproliferative syndromes (MPD) – patients, derives from the aforementioned common ancestor stem cell of HSCs and MSCs. To answer this question we assessed whether the associated with myeloid malignancy mutations and chromosomal aberrations detected in hematopoietic cells of patients, were also found in patient MSCs. We studied 16 MDS patients, 12 AML and 7 MPD, and 10 healthy age- and sex-matched individuals. DNA and RNA were isolated from the mononuclear myeloid cell fraction and cultured marrow MSCs. The presence of mutations in FLT3 gene was evaluated by genomic DNA PCR and the presence of mutations in the NPM1 gene in patients with MDS and AML was assessed by RT-PCR. The presence of mutation V617F/G1849T in JAK2 gene was detected by real time RT-PCR in patients with MPD. Moreover, MSCs and hematopoietic cells were subjected into cytogenetic analysis. In the total of MDS and AML patients, two AML patients were identified with gene mutations in bone marrow mononuclear cells: one harbored mutations in FLT3 and NPM1 genes and the other harbored a mutation in NPM1 gene. Four MPD patients were positive for the JAK2 mutation in bone marrow mononuclear cells. In respect to MSCs, none of the 35 patients harbored any of these mutations. Furthermore none of the healthy individuals harbored mutations in marrow mononuclear cells or MSCs. In 6/28 of MDS or AML patients, chromosomal abnormalities were identified in HSCs, and 1/28 in corresponding MSCs. Finally, two of healthy subjects were identified with chromosomal abnormalities in MSCs, but the corresponding HSCs were normal. In view of these data, this study gives further confirm to the absence of clonal involvement of MSCs in hematological malignancies, especially in AML, MDS and MPD.
Language Greek
Subject Acute myeloid leukemia
Chromosomal aberration
Genetic mutation
Mesenchymal stem cells
Myelodysplastic syndrome
Myeloproliferative disease
Μεσεγχυματικά κύτταρα
Μυελοδυσπλαστικό σύνδρομο
Μυελουπερπλαστικό σύνδρομο
Οξεία μυελογενής λευχαιμία
Issue date 2011-12-15
Collection   Faculty/Department--School of Medicine--Department of Medicine--Post-graduate theses
  Type of Work--Post-graduate theses
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