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Objective dermatomyositis (dermatomyositis DM) patients with normal bone marrow mesenchymal stem cells (bone marrow-derived mesenchymal stem cells, BMSCs) biological traits, to explore the the DM patients BMSCs whether defective or variation, whether applied to autologous transplant, or DM itself is a stem cell disease, associated with the incidence of DM, which lay the foundation for the clinical application of autologous transplantation in patients with DM BMSCs. Method 1. Were collected 5 cases of DM patients with normal bone marrow each 5 ml of-8ml cells were isolated by density gradient centrifugation and adherent culture, subculture, FACS surface marker identification of P3 generation of BMSCs. BMSCs proliferation capacity comparison of the two groups: MTT method determination of P3, P6 generation BMSCs growth curve, calculate the cell population doubling time; flow cytometry to determine the cell cycle, calculate the proliferation index; real-time quantitative PCR (real time Polymerase chain reaction , RT-PCR) determination of the expression of human telomerase reverse transcriptase (human telomerase reverse transcriptase, hTERT). Two groups BMSCs comparison to the ability of vascular endothelial cells: the P3 generation of cells induced in endothelial cells culture medium (H-DMEM culture medium with 10% fetal bovine serum 100u/ml penicillin 100pg/ml streptomycin 10ng/ml VEGF 5ng/ml bFGF) in the culture, and L-DMEM culture solution cultured normal human BMSCs as a negative control, observed under inverted microscope cell growth and morphology, expression of factor Ⅷ 14d immunohistochemistry, flow cytometry CD34, vascular endothelial growth factor receptor (kinase-insert domain conteining receptor, KDR) expression of the case, so as to identify the of BMSCs immunogenicity endothelial cells. Results of the two groups can be successfully isolated and cultured of BMSCs P3 generation no significant differences in cell morphology, adherent cells were highly homogeneous, long spindle or polygonal, parallel to, swirly, radial order. By flow cytometry, CD34, CD45-negative, CD44-positive, CD71 weakly positive, identified as mesenchymal stem cells. BMSCs proliferation capacity comparison of the two groups: normal control group P3 generation cell population doubling time (4.73 ± 0.31) d, DM group (4.76 ± 0.32), no statistically significant difference between the two groups, P gt; normal control group 0.05; P6 generation cell population doubling time (5.74 ± 0.35) d DM group (6.14 ± 0.34) d, the DM group BMSCs population doubling time longer than the control group, the difference was statistically significance (P lt; 0.001 ). P3 generation BMSCs proliferation index of the normal control group (17.87 ± 2.05)% DM group (17.12 ± 2.00)%, the difference was not statistically significant (P gt; 0.05); the P6 generation normal control group BMSCs proliferation index ( 10.73 ± 1.78)%, DM group (5.96 ± 2.16)% in the DM group proliferation index is less than the control group, a statistically significant difference (P lt; 0.01). Normal control group P6 generation BMSCs hTERT expression (10.63 ± 1.49), DM group (6.45 ± 2.04), DM patients with hTERT expression levels compared with normal control group, the difference was statistically significant (P lt; 0.01). Two groups of cells in culture 14d Immunohistochemical detection of the expression of factor Ⅷ induced positive flow cytometry KDR, CD34 expression was positive cells of the DM group and the control group compared to the expression of CD34, the difference was not statistically significant (P gt; 0.05). DM group compared with the control group cells expressing KDR less, the difference was statistically significant (P lt; 0.05). This may be related to small sample size, may also be due to the DM itself BMSCs abnormality. Conclusions of the two groups have successfully isolated and cultured of BMSCs, and the DM no significant difference between patients and the normal control group cell morphology; 2 DM patients BMSCs flawed in varying degrees; 3 between the two groups BMSCs in vitro can be induced into class vascular endothelial cells, the induction class vascular endothelial cells have some differences; DM patients BMSCs for autologous transplantation, solutions can be considered allogeneic BMSCs transplantation.
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