RESUMEN
Objective To observe the effect of fluvastatin(Flu) on differentiation and induction of human promyelocytic leukemia HL-60 cells and provide the theoretical foundation for treatment of human promyelocytic leukemia.Methods The cultured HL-60 cells were treated with 20 ?mol?L-1 Flu,the morphological changes of the cell differentiation were examined.The NBT reduction capability was detected in HL-60 cells treated with Flu for 72 h.After HL-60 cells were treated with 20 ?mol?L-1 Flu for 5 d,they were stained with non-specific esterase and the percentage of differentiation cells was analyzed.Results The HL-60 cells treated with Flu showed smaller cell body,reducing proportion of nucleus to cytoplasm,the nucleus tortuosity,fold or sublobe.There were specific granules and vacuoles in cytoplasm,displaying that some cells had differentiated to relative mature cells.As compared with control group,the NBT reduction capability in HL-60 cells treated with Flu for 72 h was significantly higher than that in control cells(P
RESUMEN
Objective:To establish the technique for T lymphocytes inducted and differentiated by using human hematopoietic stem/progenitor cells ex vivo and to provide a technological platform for studying T cells biological characteristics and cellular immune.Methods:Human CD34-positive cells were isolated from umbilical cord blood by using a high-gradient magnetic cell sorting system(MACS) and CD34+ cells were seeded on human fetal thymic stromal monolayer system.Liquid culture system contains hematopoietic cytokines(FL+IL-12+IL-7+IL-2) and 20% human AB serum.Non-adhension cells were obtained after 7?14?28?35?42 days of culture respectively to analysis T cells phenotype by FACS using T cells specific monoclonal antibody.T cells morphology were also studied.Results:After 2 weeks of culture CD4+CD8+ immature T lymphocytes wre about 0 3%～13 3% and the peak value is 16 6%～26 5% at 4～5 weeks of culture.CD4-CD8+ and CD4+CD8- T cells that coexpressed CD3 cells increased at 26 5%～64 9% and 11 6%～38 9% after 6 weeks of culture.Wright staining shows that the mature T cells harvested after mitogenic stimulation have the presence of large cells with lymphoblast morphology.Conclusion:Human umbilical cord blood CD34+ cells could be differentiated into T lymphocytes by the culture conditions of human fetal thymic stromal monolayer system and the cytokine combination of FL+IL-12+IL-7+IL-2,and the T cells can be stimulated by IL-2+PHA.
RESUMEN
Purpose:To study the capacity of hematopoietic growth factors combinations(IL 3,G CSF,GM CSF) for inducing CD34 + cells to proliferate and differentiate committedly to granulocytes. Methods:CD34 + cells were isolated from peripheral blood by using a high gradient magnetic cell sorting sysem(MACS),and expanded with IL 3、G CSF and GM CSF in a liquid culture system.Colony forming cells and CD34 + cell ratio were studied by colony forming assays and FACS. Results:The combinations of IL 3,G CSF and GM CSF increased the total cell number by 56 fold and 110 fold, increased CFU GM by 8.44 fold and 35.08 fold respectively at the 3rd and the 7th day after culturing ,and the ratio of CD34 +cell decreased during the culture.Conclusions:It is possible to induce CD34 +cells to proliferate and differentiate committedly to granulocyte lineage.
RESUMEN
Inhibition of proliferation and induction of differentiation by 1, 25-dihydroxyvitamm D3 of human promyelocytic leukemic cell line (HL-60)are reported. At the concentration range of 109 to 106 mol/ L, 1, 25-dihydroxyvitamin D3 had effect in inhibiting cell proliferation with time-dependent and dose-dependent manner. Strong inhibition of colony cell growth (74.8?11.1%)was observed at 107 mol/L concentration. It was also found that HL-60 cells could be induced into monocytes by 1, 25-dihydroxyvitamin D3at the same concentrations,which were demonstrated by cytochemistry and specific antigen of cell surface and NBT reduction test. In addition, the relationship between proliferation and differentiation is discussed according to the result of flow cytometry.