Publication Date:
2007-11-16
Description:
Myelodysplastic syndromes (MDS) are a group of clonal disorders of hematopoietic stem cell (HSC). The hematopoietic microenvironment plays a major role in the physiology of the hematopoietic system, and mesenchymal stem cells (MSC) are not only the progenitors but also the key regulators of this microenviroment. Recently, some data has been published showing that these MSC could be involved in the MDS pathophysiology. Moreover, the presence of cytogenetic aberrations on these cells is controversial. The aim of the study was to characterize bone marrow derived MSC from patients with MDS using different approaches: kinetic studies, immunophenotypic analysis and genetic changes by array-based comparative genomic hybridization (array-CGH). FISH was performed with the probe of 1q31 and Q-PCR was performed with the SYBR Green technique in order to confirm array-CGH results. Patients with untreated MDS were included in the study. Median age was 72 years (range: 54–89). Diagnosis of MDS was established according to the WHO classification as follows: 5q- syndrome (n=7), refractory anemia (n=2), refractory anemia with ringed sideroblasts (n=1) and refractory anemia with excess blasts type 2 (n=3). Standard cytogenetic and FISH studies on hematopoietic cells were performed at diagnosis according to standard methods. MSC from MDS were compared to those from 12 healthy donors. MSC were obtained by plating mononuclear cells from bone marrow, and cultured and expanded following standard procedures. According to the international consensus for MSC characterization, in the third passage MSC were harvested to perform phenotypical studies and cytogenetics. To perform Array-CGH a total of 3500 genomic targets were compounded from RP-11 libraries. The PCR products after purification were arrayed onto glass slides using a BioRobot. DNA was labelled, denaturalised and hybridizated. MSC from MDS achieved confluence at a slower rate than donor-MSC [23 days (range 12–90) vs 15 days (range 11–30) p
Print ISSN:
0006-4971
Electronic ISSN:
1528-0020
Topics:
Biology
,
Medicine
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