A Study Of The Biological Character Of DM Patients' BMSCs

Objective By comparing the biological characteristics of bone marrow mesenchymal stem cells (BMSCs) between the dermatomyositis (DM) patients and normal human to discuss that whether the defects or variation exist on the BMSCs of DM patients, whether self-transplant can be applied in DM patients, or DM itself is a stem cell disease, and to build the theoretiacal foundation of the clinical application of BMSCs transplant.Methods 5-8ml bone marrow of five DM patients and five normal control volunteers were collected respectively. The cells were isolated culture by gradient centrifugation and adherence cultivation, and the surface markers of BMSCs were identitied by flow cytometer. Firstly BMSCs'reproductive activity of the groups were compared: the auxodrome and cell cycle of BMSCs were maeasured respectively, and the doubling generation time of cell colony and proliferation index were calculated in order to see whether there was a dufference of the reproductive activity between the two groups. Further more the lengeth of perultimate chromomere and hTERT were measured. Besides, capability of cell differentiation of the two groups were compared: the third generation(P3) cells were cultured in endotheliocyte induction culture solution and L-DMEM culture fluid separately. Observate the growth and morphous under inverted microscope. To identify the endotheliocyte stem from MSCs, facterâ…§was detected by immunohistochemi stry. Kinase-insert domain conteining receptor (KDR), CD34 was detected by flow cytometry.Results The results of flow cytometry showed that CD34 and CD45 were negative, CD44 was positive, and CD71 was lightly positive. the morphology of the two group cells were similar after adherence, long fusiform, polygon, parallel, vortex-shaped or radially arranged. Between the two groups, the difference of the cell colony doubling generation time of and proliferation index had no the statistical significance at P3, but had the statistical significance at P6. Determination of hTERT among the two groups had statistical significance. After induced 14 days, the results of immunohistochemistry of two cells showed facterâ…§was positive, the results of flow cytometry showed that KDR and CD34 were positive. DM group and control group compared with CD34 cells, the difference was not statistically significant. DM group compared with the control group cells to express KDR less, the difference was statistically significant. This may be less concerned with the sample size , but also may be due to abnormal MSCs of DM itself.Conclusion Two groups can be successfully isolated and cultured to obtain BMSCs, and DM patients and normal control has no significant differences in cell morphology. DM group compared with the control group cells to express KDR less, the difference was statistically significant. It is speculated, the existence of DM patients with BMSCs defects. So allogeneic BMSCs transplanted should be a choose for cell therapy.