The Primary Research On The Abnormality Of Marrow Mesenchymal Stem Cells From Patients With Bone Marrow Failure Syndrome

ObjectiveTo investigate the differences in morphology and growth characteristics of mesenchymal stem cells (MSCs) derived from bone marrow of aplastic anemia (AA) patients and myelodysplastic syndrome (MDS) patients, and to research cytokines mRNA expressions on bone marrow MSCs from these patients, as well as to explore the role of these abnormalitis in the pathogenesis of AA and MDS.MethodsMSCs were isolated from bone marrow sample of AA patients, MDS patients and normal individuals and cultured in vitro.The morphology and growth cures feature were observed. The semi-quantitiative reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect mRNA the expressions of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), interleukin-6 (IL-6), IL-1β, stem cell factor (SCF), granulocyte colony stimulating factor (G-CSF) at mRNA level for bone marrow MSCs of patients with AA and MDS. The real time quantitative polymerase chain reaction (RQ-PCR) technique was used to detect the mRNA expression of thrombopoietin (TPO) in bone marrow MSCs from AA and MDS patients. ResultsMSCs derived from AA patients and normal individuals were similar in cell morphology. Both of cells was fibroblast-like spindle. But MSCs from AA patients grew slower than of the normal control group. The time of colony-forming of the MSCs from AA patients was longer.And the time required for the MSCs from AA patients reaching 90% confluency was also longer than the normal control group. The proliferation capacity of MSCs from AA patients at passage 3 was weaker than that of normal individuals. While, MDS patients and normal individuals derived MSCs were similar in cell morphology as well as the time of cell colony-forming and cell reaching 90% confluency. There was no difference in the proliferative ability of MSCs at passage 3 between MDS patients and normal control group. Compared with normal control group, the expressions of VCAM-1 and ICAM-1 of MSCs of AA had no significant difference (P>0.05). The expressions of IL-6 and SCF of them were much lower than that of the normal group (P<0.05), and the expression of TPO of them was higher than that of the normal (P<0.05), while the expression of IL-1βof them had no significant difference when contrasted to the normal (P<0.05). The expressions of VCAM-1 and ICAM-1 of MSCs of MDS had no significant difference when contrasted to the normal group (P>0.05). Compared with normal control group, the expression of SCF of them was lower (P<0.05), but the expressions of IL-6,IL-1βand TPO of them showed no significant difference (P<0.05). The expression IL-6 of MSCs of AA was significantly lower than that of the MDS (P<0.05), but for the expressions of VCAM-1, ICAM-1, IL-1β, SCF and TPO, there were no significant difference between two groups (P>0.05). Neither the patients nor the normal group had the expression of G-CSF.Conclusion The morphology of MSCs showed a similar characteristics among the three groups. But compared with MSCs from normal individuals, MSCs from AA showed significant slower growth rate and weaker proliferative ability. Whereas, there were no significant difference between MSCs of MDS and that of normal control group in morphology, growth rate and proliferation capacity. The expressions of VCAM-1 and ICAM-1 were no difference in bone marrow MSCs of AA and MDS. The abnormalities of the expressions of IL-6,SCF and TPO of bone marrow MSCs in AA patients were positive and the expression of SCF is abnormal in bone marrow MSCs from MDS patients. All of thses comfirm the bome marrow MSCs of AA and MDS are dysfunction.