The Minimal Residual Disease And The Morphology Of CD34 Positive Hematopoietic Cells

The first chapter The minimal residual disease of CD34 positive hematopoietic cellsObjective:To explore the sensitivity of fluorescence in situ hubridization (FISH) for the minimal residual disease (MRD) of CD34 positive cells. Materials and Methods:The samples of bone marrow were taken from 10 CD34 positive leukemia patients who had been complete remission. CD34 positive cells were purified by MiniMACS and detected by the flow cytometer. The two groups of samples included bone marrow and CD34 positive cells were detected by FISH. The data of two groups were analyzed by paired Mest. Results:The average purity of CD34+ cells in mononucleic cells before purify was (0.7?.2)%,and the average purity of CD34+ cells after purify was (90.4?.4)%. There were average 1.8% and 4 in 10 positive results before purify and average 4.7% and 9 in 10 positive results after purify. There were significant difference between two groups, P equal 0.037. Conclusion:Associating the magnetic cell sorting and FISH, the sensitivity of FISH which detect for the minimal residual disease can be improved significantly.The second chapter The morphology of CD34 positive hematopoietic ceUsObjective:To explore and compare the morphology of CD34+ cells of different groups which include the bone marrow and peripheral blood of health human, the bone marrows of patients with AML (M2b), the peripheral blood of patients with complete remission leukemia and the SLE by optical and atomic forcemicroscopes (AFM). Materials and Methods:CD34 positive cells were purified by MACS and detected by the flow cytometer. The CD34+ cells of the bone marrow and peripheral blood of health human and the bone marrow of patients with AML (M2b) were observed by optical microscope with May-Grunwald-Giemsa staining. The CD34+ cells from the bone marrow of a patient with AML (M2b) and a health human were made of two blocks of resin. Then ultrathin slices of cells about 60 nanometer were made by ultramicrotome. These ultrathin slices of cells were observed by atomic force microscopes respectively. Atomic force microscope also was used to observed the surface of the CD34+ cells of each group. Twenty cells were observed randomly in each group. The average of some parameters such as mean height and roughness of five images in each cell represented as the whole cell. The data of five groups were analyzed by One Way ANOVA. Results:The average purity of CD34+ cells in MNC was (1.63+0.25) %, and the average purity of CD34+ cells after purify by MiniMACS was (91+2.4) %, and the average purity of CD34+ cells after purify by ClinMACS was (95+2.7) %. The results of the optical microscope suggested that there were no significant difference between the CD34 +cells of the bone marrows and peripheral blood of health human. There were significant difference between the CD34+cells of the bone marrow of patients with AML (M2b) and the others. AFM could be used to observe the structure of the inner CD34+cells. The results suggested that there were no significant difference between the patient with AML (M2b) and health human from the bone marrow. When the scope of images were 10 X 10 u m, the nucleolus and some organelles such as mitochondria could be identified by AFM. When the scope of images were 2X2 u m, the images were obscure because of many tiny granular resin. There were many rough granules and erosive structure on the surface of the CD34+ cells observed by AFM. When the scope of images were 10 X 10 u m, the whole cell could be imaged. When the scope of images were 2 X 2 u m, there were more rough and erosive structure on the surface of the CD34+ cells. There weresignificant difference in respects of some parameters such as mean height and roughness between the sort of CD34+ cells from the bone marrow of AML (M2b) patients with the other sorts of CD34+ cells and there were no significant difference between the other sorts of CD34+ cells. Conclusion:It can be concluded that the CD34+ cells can be purified by magnetic cell sorting to match the study of morphology.