To clarify the hematopoietic potential of various sub-classes of human hematopoietic progenitor cells, we used a multicolor staining protocol in conjunction with anti-CD34 and -CD38 McAb. We characterized two cell fra...To clarify the hematopoietic potential of various sub-classes of human hematopoietic progenitor cells, we used a multicolor staining protocol in conjunction with anti-CD34 and -CD38 McAb. We characterized two cell fractions in CD34+cells with or without CD38 expression. A clonogenic assay showed that most CFC were present in CD34+CD38+ population. Morphologic analysis showed that blast-like cells were more enriched in the CD34+CD38 fraction. To clarify the biologic differences between both fractions, we examined the more primitive progenitor cell function by assessing long-term culture-initiating cells (LTC-IC) on the stromal cells. At the first two weeks, more CF.C harvested from the culture in the fractions initiated with both populations. However, more LTC-IC were present during weeks 4 to 12 in the CD34+CD38- population. These results indicate the primitive progenitors are more enriched in CD34+CD38 population than in CD34+CD38+ cells.展开更多
AIM To evaluate the importance of the CD34+CD38-cell population when compared to the CD34+CD38+/low and CD34+CD38+/high leukemic cell sub-populations and to determine its correlations with leukemia characteristics and...AIM To evaluate the importance of the CD34+CD38-cell population when compared to the CD34+CD38+/low and CD34+CD38+/high leukemic cell sub-populations and to determine its correlations with leukemia characteristics and known prognostic factors, as well as with response to therapy and survival.METHODS Two hundred bone marrow samples were obtained at diagnosis from 200 consecutive patients with newly diagnosed acute myeloid leukemia(AML) were studied between September 2008 and December 2010 at our Institution(Hematology Department, Lyon, France). The CD34/CD38 cell profile was analyzed by multiparameter flowcytometry approach using 8 C panels and FACS CANTO and Diva software(BD Bioscience).RESULTS We analyzed CD34 and CD38 expression in bone marrow samples of 200 AML patients at diagnosis, and investigated the prognostic value of the most immature CD34+CD38-population. Using a cut-off value of 1% of CD34+CD38-from total "bulk leukemic cells" we found that a high(> 1%) level of CD34+CD38-blasts at diagnosis was correlated with advanced age, adverse cytogenetics as well as with a lower rate of complete response after induction and shorter disease-free survival. In a multivariate analysis considering age, leukocytosis, the % of CD34+ blasts cells and the standardized cytogenetic and molecular risk subgroups, a percentage of CD34+CD38-leukemic cells > 1% was an independent predictor of DFS [HR = 2.8(1.02-7.73), P = 0.04] and OS [HR = 2.65(1.09-6.43), P = 0.03].CONCLUSION Taken together, these results show that a CD34/CD38 "backbone" for leukemic cell analysis by multicolour flowcytometry at diagnosis provides useful prognostic information.展开更多
BACKGROUND To date,no specific treatment has been established to reverse progressive chronic kidney disease(CKD).AIM To evaluate the safety and efficacy of autologous CD34^(+)cell transplantation in CKD patients who e...BACKGROUND To date,no specific treatment has been established to reverse progressive chronic kidney disease(CKD).AIM To evaluate the safety and efficacy of autologous CD34^(+)cell transplantation in CKD patients who exhibited a progressive decline in renal function.METHODS The estimated glomerular filtration rate(eGFR)at the beginning of the study was 15.0-28.0 mL/minute/1.73 m^(2).After five days of treatment with the granulocyte colony-stimulating factor,mononuclear cells were harvested and CD34^(+)cells were magnetically collected.CD34^(+)cells were directly injected into the bilateral renal arteries twice(at 0 and 3 months),and their safety and efficacy were evaluated for 6 months.RESULTS Four patients were enrolled and completed the study.Three of four patients showed improvement in eGFR slope(eGFR slope>0 mL/minute/1.73 m^(2)),with the monthly slope of eGFR(delta eGFR)changing from-1.36±1.1(pretreatment)to^(+)0.22±0.71(at 6 months)mL/minute/1.73 m^(2)/month(P=0.135)after cell therapy.Additionally,intrarenal resistive index(P=0.004)and shear wave velocity(P=0.04)were significantly improved after cell therapy.One patient experienced transient fever after cell therapy,and experienced bone pain during granulocyte colony-stimulating factor administration.However,no severe adverse events were reported.CONCLUSION In conclusion,our findings suggest that repetitive peripheral blood-derived autologous CD34^(+)cell transplantation into the renal arteries is safe,feasible,and may be effective for patients with progressive CKD.However,a large-scale clinical trial is warranted to validate the efficacy of repetitive regenerative cell therapy using autologous CD34^(+)cells in patients with progressive CKD.展开更多
CD34+ cells from human umbilical cord blood were purified by Dynal beads M-450 CD34 immunoselection system and cultured in the presence of various cytokines alone or in combination, including stem cell factor (SCF), i...CD34+ cells from human umbilical cord blood were purified by Dynal beads M-450 CD34 immunoselection system and cultured in the presence of various cytokines alone or in combination, including stem cell factor (SCF), interleukin-6 (IL-6) and erythropoietin (EPO). The results revealed that: (D In methylcellulose culture, the plating efficiencies of purified cord blood CD34+ cells were much different when stimulated by various cytokines. IL-6 alone had the lowest colo-ny yield, while the combination of SCF, IL-6 and EPO had the highest yield. ② In the suspension culture, IL-6 alone or IL-6 + EPO had little expanding effect on cord blood CD34+ celis, the other cytokine combinations could expand cord blood CD34+ celis at different Ievels. Among them, the combination of SCF, IL-6 and EPO had the maximal expanding effect on cord blood CD34+ celis, the number of progenitor celis peaked at day 21, about 29-fold increase and nucleated celis increased approximately 3676-fold at day 28. The expanding effect of展开更多
文摘To clarify the hematopoietic potential of various sub-classes of human hematopoietic progenitor cells, we used a multicolor staining protocol in conjunction with anti-CD34 and -CD38 McAb. We characterized two cell fractions in CD34+cells with or without CD38 expression. A clonogenic assay showed that most CFC were present in CD34+CD38+ population. Morphologic analysis showed that blast-like cells were more enriched in the CD34+CD38 fraction. To clarify the biologic differences between both fractions, we examined the more primitive progenitor cell function by assessing long-term culture-initiating cells (LTC-IC) on the stromal cells. At the first two weeks, more CF.C harvested from the culture in the fractions initiated with both populations. However, more LTC-IC were present during weeks 4 to 12 in the CD34+CD38- population. These results indicate the primitive progenitors are more enriched in CD34+CD38 population than in CD34+CD38+ cells.
文摘AIM To evaluate the importance of the CD34+CD38-cell population when compared to the CD34+CD38+/low and CD34+CD38+/high leukemic cell sub-populations and to determine its correlations with leukemia characteristics and known prognostic factors, as well as with response to therapy and survival.METHODS Two hundred bone marrow samples were obtained at diagnosis from 200 consecutive patients with newly diagnosed acute myeloid leukemia(AML) were studied between September 2008 and December 2010 at our Institution(Hematology Department, Lyon, France). The CD34/CD38 cell profile was analyzed by multiparameter flowcytometry approach using 8 C panels and FACS CANTO and Diva software(BD Bioscience).RESULTS We analyzed CD34 and CD38 expression in bone marrow samples of 200 AML patients at diagnosis, and investigated the prognostic value of the most immature CD34+CD38-population. Using a cut-off value of 1% of CD34+CD38-from total "bulk leukemic cells" we found that a high(> 1%) level of CD34+CD38-blasts at diagnosis was correlated with advanced age, adverse cytogenetics as well as with a lower rate of complete response after induction and shorter disease-free survival. In a multivariate analysis considering age, leukocytosis, the % of CD34+ blasts cells and the standardized cytogenetic and molecular risk subgroups, a percentage of CD34+CD38-leukemic cells > 1% was an independent predictor of DFS [HR = 2.8(1.02-7.73), P = 0.04] and OS [HR = 2.65(1.09-6.43), P = 0.03].CONCLUSION Taken together, these results show that a CD34/CD38 "backbone" for leukemic cell analysis by multicolour flowcytometry at diagnosis provides useful prognostic information.
文摘BACKGROUND To date,no specific treatment has been established to reverse progressive chronic kidney disease(CKD).AIM To evaluate the safety and efficacy of autologous CD34^(+)cell transplantation in CKD patients who exhibited a progressive decline in renal function.METHODS The estimated glomerular filtration rate(eGFR)at the beginning of the study was 15.0-28.0 mL/minute/1.73 m^(2).After five days of treatment with the granulocyte colony-stimulating factor,mononuclear cells were harvested and CD34^(+)cells were magnetically collected.CD34^(+)cells were directly injected into the bilateral renal arteries twice(at 0 and 3 months),and their safety and efficacy were evaluated for 6 months.RESULTS Four patients were enrolled and completed the study.Three of four patients showed improvement in eGFR slope(eGFR slope>0 mL/minute/1.73 m^(2)),with the monthly slope of eGFR(delta eGFR)changing from-1.36±1.1(pretreatment)to^(+)0.22±0.71(at 6 months)mL/minute/1.73 m^(2)/month(P=0.135)after cell therapy.Additionally,intrarenal resistive index(P=0.004)and shear wave velocity(P=0.04)were significantly improved after cell therapy.One patient experienced transient fever after cell therapy,and experienced bone pain during granulocyte colony-stimulating factor administration.However,no severe adverse events were reported.CONCLUSION In conclusion,our findings suggest that repetitive peripheral blood-derived autologous CD34^(+)cell transplantation into the renal arteries is safe,feasible,and may be effective for patients with progressive CKD.However,a large-scale clinical trial is warranted to validate the efficacy of repetitive regenerative cell therapy using autologous CD34^(+)cells in patients with progressive CKD.
文摘CD34+ cells from human umbilical cord blood were purified by Dynal beads M-450 CD34 immunoselection system and cultured in the presence of various cytokines alone or in combination, including stem cell factor (SCF), interleukin-6 (IL-6) and erythropoietin (EPO). The results revealed that: (D In methylcellulose culture, the plating efficiencies of purified cord blood CD34+ cells were much different when stimulated by various cytokines. IL-6 alone had the lowest colo-ny yield, while the combination of SCF, IL-6 and EPO had the highest yield. ② In the suspension culture, IL-6 alone or IL-6 + EPO had little expanding effect on cord blood CD34+ celis, the other cytokine combinations could expand cord blood CD34+ celis at different Ievels. Among them, the combination of SCF, IL-6 and EPO had the maximal expanding effect on cord blood CD34+ celis, the number of progenitor celis peaked at day 21, about 29-fold increase and nucleated celis increased approximately 3676-fold at day 28. The expanding effect of