| Objective: To investigate the origination of abnormal clonal cells in bone marrow of patients with MDS and to study the condition of clonal cells that is derived from CD34~+CD38~- and CD34~+CD38~+ cells after long-term culture.Methods: Clonal cell proportions in CD34~+CD38~- and CD34~+CD38~+ cells from bone marrow of MDS patients were detected. The cytospins of BM karyotypes were got by hemalysis and Immuno-Magnetic Bead technique was used to sort CD34~+CD38~- and CD34~+CD38~+ cells from bone marrow monocular cells of eleven MDS patients with chromosome abnormalities(four cases with trisomy 8, complex karyotype with trisomy 8 in two case, two cases with 5q-, complex karyotype with 5q- in two case,5q- accompanying trisomy 8 in one case) and got smears respectively. The percentage of abnormal clones in CD34~+CD38~- and CD34~+CD38~+ cells were compared by fluorescent in sire hybridization (FISH). Furthermore, CD34~+CD38~- and CD34~+CD38~+ cells of two patients with +8 were added to long-term culture mediums respectively for 6 weeks and plated in complete methylcellulose cultures to observe colony-formation after 14 days.Results: 1,abnormal clonal cells were detected in both CD34~+CD38~- and CD34~+CD38~+ cells from eleven patients with 5q- and +8. The percentage of abnormal clones in CD34~+CD38~- cells (46.3±8.1)% were obviously lower than those in CD34~+CD38~+ cells (75.9±6.8)% , p<0.001, and there was no obviously difference on the percentage of abnormal clones between CD34~+CD38~+ cells and bone marrow karyocytes,p=0.833;2,CD34~+CD38~- and CD34~+CD38~+ cells from the two patients with +8 were cultured and no colonies were produced.Conclusion: It can be suggested that abnormal clones in bone marrow originated from stem cells in MDS patients with 5q- and +8 and the abnormal clones may be prior to normal hematopoietic cells in progenitors'level and may conclude that apoptosis increased in abnormal clones during late stage of differentiation. |
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