Effects of Astragalus-Angelica compatilibity on bone marrow hematopoiesis suppression induced by Cyclophosphamide in mice / 中草药

Objective: To investigate the effects of different proportion combinations of Astragalus and Angelica on bone marrow hematopoiesis suppression induced by Cyclophosphamide (CTX) in mice, and to analysis their interactions. Methods: The model of bone marrow hematopoietic function suppression in mice was established by ip injection of CTX. Recombinant human granulocyte-macrophage colony-stimulating factor (rhG-CSF) was used as the positive control drug, ICR mice in the drug groups were administered with the extracts of Astragalus, Angelica, and combinations of Astragalus and Angelica with different ratios (10:1, 5:1, 2.5:1, 1:1, 1:2.5, 1:5, and 1:10). To detect peripheral hemogram, the nucleated cell count in bone marrow (BMNC), the area of bone marrow hematopoietic tissue, spleen index (SI), as well as the contents of hematopoietic growth factor (HGF) in serum such as Erythropoietin (EPO), thrombopoietin (TPO), and Granulocyte-macrophage colony-stimulating factor (GM-CSF). To integrate all the test parameters using comprehensive index method, then to analyze the interaction between Astragalus and Angelica through multiple linear regression analysis. Results: Following continuous CTX injection for 3 d, peripheral hemogram significantly decreased, accompanied by GM-CSF and TPO contents declined, BMNC reduced, SI elevated, and bone marrow hematopoietic tissue area lessened from day 5 to day 7 after injection. Single Astragalus had no remarkable effects on peripheral hemogram, HGF contents, bone marrow hematopoietic tissue area and BMNC. Except for HGF and BMNC, Angelica alone could increase the numbers of WBC, RBC, and PLT in the peripheral blood, raise the area of bone marrow hematopoietic tissue. Astragalus mixed Angelica with the ratios of 5:1, 2.5:1, 1:1, 1:2.5, 1:5, and 1:10 could increase the numbers of WBC, RBC, and PLT along with HGF content, BMNC and bone marrow hematopoietic tissue area, while decreased SI. The comprehensive effect analysis displayed that the effects of Astragalus-Angelica with 1:1, 1:2.5, and 1:5 ratios on promoting hematopoiesis were strongest. The interaction analysis revealed Angelica played a greater role in advancing hematopoiesis than Astragalus did after being combined, meanwhile, the interaction of Astragalus combined with Angelica produced a nonnegligible positive effect on promoting hematopoiesis, namely synergism. Conclusion: Astragalus-Angelica compatilibity has the synergism on promoting hematopoiesis, which is better when the combination ratio of Astragalus and Angelica is 1:1, 1:2.5, or 1:5, furthermore, the effect of Angelica is greater than that of Astragalus.

Clinical application of cytokines in acute iatrogenic radiation damage / 中华放射医学与防护杂志

Objective To explore the influence of hematopoietic growth factors on the TBI patients in the conditioning regimen for stem cell transplantation,and to evaluate the effect of cytokines on treatment of acute radiation disease.Methods The usage of hematopoietic growth factors,implantations and the side-effects of the patients and donors of TBI in the conditioning regimen for stem cell transplantation were retrospectively analysed from 1990 to 2012,and the effect and side-effects of cytokines on the hematopoietic function recovery and stem cell mobilization were observed.Results All patients recovered from their hematopoietic function except one died due to the side-effecs.The median time of white blood cell recover was 10 d in auto-SCT and 12 d in allo-SCT in the G-CSF group.The median day of platelet recovery was 11.67 ± 1.53 in rhIL-11 arm and 13.70 ±6.27 in no rhIL-11 arm in the auto-SCT group.The incidence rates of dental ulcer and diarrhea in the TBI patients were 48％ and 44％,respectively.The occurrence of side-effect was rare in the period of cytokines treatment,but was over 50％ during stem cell mobilization.Conclusions Cytokines play very important roles in the hematopoietic function recovery and stem cell mobilization in the TBI patients.

Effects of Short-Term Incubation of Hematopoietic Cells with Hematopoietic Growth Factors on CXCR4 Expression and Stromal Cell-Derived Factor-1-Mediated Transendothelial Migration / 대한혈액학회지

BACKGROUND: There has been contradictory reports regarding the homing potential of hematopoietic cells briefly exposed to hematopoietic growth factors in vitro. To get a resolution to this controversy, we investigated the effects of short-term growth factor treatment of hematopoietic cells on the expression of CXCR4 and adhesion molecules, and the chemotaxis in response to stromal cell-derived factor-1 (SDF-1), which is widely accepted to play a critical role in bone marrow (BM) homing of hematopoietic stem cells. METHODS: BM and cord blood(CB) CD34+ cells were incubated with various hematopoietic growth factors including IL-1beta, IL-3, IL-6, G-CSF, GM-CSF, stem cell factor (SCF), flk-2 ligand, and thrombopoietin, alone or in combination for up to 48 hours. Before and after the incubation, the expression of CXCR4 and adhesion molecules of CD34+ cells was analyzed using flow cytometry. SDF-1-mediated transmembrane or transendothelial migration of CD34+ cells, cobblestone area-forming cells (CAFCs), and/or long-term culture-initiating cells (LTC-ICs) was measured using Transwell(TM) system. RESULTS: VLA-4 was moderately up-regulated by the incubation of the cells with IL-3 and SCF, and ICAM-1 was slightly up-regulated by IL-1 and IL-3. The expression of L-selectin, PECAM-1 or LFA-1 was not altered by any growth factors. With the incubation of the cells in the absence of growth factors or SDF-1, CXCR4 expression of CD34+ cells was rapidly increased, reaching a plateau at 24 hours. The spontaneous up-regulation was abrogated with the addition of SDF-1. In agreement with the up-regulation of CXCR4, CD34+ cells incubated for 40 hours showed much enhanced chemotaxis in response to SDF-1 compared to non-incubated cells (24.7 3.5% vs. 7.0 1.6%, P=0.01). Any growth factors examined in this study did not alter the CXCR4 expression of CD34+ cells. Neither did growth factors affect the transendothelial migration of LTC-ICs toward bone marrow stromal cells as well as the SDF-1-induced transmembrane chemotaxis of CD34+ cells and CAFCs. CONCLUSION: Short-term treatment of hemo-topoietic cells with hematopoietic growth factors does not alter the expression of CXCR4 or SDF-1-mediated transendothelial chemotaxis.

Ex Vivo Expansion of Cryopreserved Cord Blood Cells / 대한혈액학회지

BACKGROUND: Cord blood (CB), which has no HLA restriction, is an alternative to bone marrow for hematopoietic stem cell transplantation. The use of cord blood, however, is limited by the number of progenitor/stem cells necessary to reconstitute the older child or adult. Therefore, ex vivo expansion of CB could have tremendous impact on diverse clinical settings. We studied the ex vivo expansion of isolated population of CD34+ cells from cryopreserved CB cells. METHODS: CD34+ cells were isolated from cryopreserved CB mononuclear cells. Purified cells were cultured with various combinations of hematopoietic growth factors including erythropoietin (EPO), stem cell factor (SCF), granulocyte-colony-stimulating factor (G-CSF), granulocyte, macrophage-colony-stimulating factor (GM-CSF), interleukin-1beta (IL-1beta), IL-3, and IL-6. After 7, 10 or 14 days of culture, the fold increases of colony-forming unit- granulocyte, macrophage (CFU-GM), burst-forming unit-erythroid (BFU-E), colony-forming unit-mix (CFU-Mix), and high proliferative potential colony-forming cell (HPP-CFC) were evaluated. RESULTS: Ten-day culture with the combination of EPO, SCF, G-CSF, IL-1beta, and IL-3 resulted in a median of 60-fold increase of CFU-GM, which was greater than those with the combinations of less than 5 growth factors. The addition of IL-6 or GM-CSF to this combination did not enhance CFU-GM expansion. Ten-day culture was significantly superior to 7-day culture for CFU-GM expansion. Prolongation of culture to 14 days, however, revealed decreased expansion of CFU-GM compared to 10 days. BFU-E and CFU-Mix were expanded to 2~5 folds in 7-day culture with the combination of EPO, SCF, and G-CSF. Further expansion was not achieved in 10-day culture and colonies disappeared in 14-day culture. HPP-CFC was expanded to a median of 7.5 folds in 7-day culture with the combination of EPO, SCF, G-CSF, IL-1beta, IL-3, and IL-6. Neither 10-day or 14 day-culture enhanced expansion of HPP-CFU. CONCLUSION: Cryopreserved cord blood cells maintain ex vivo expansion potential. In our system, 10-day culture with the combination consisting of EPO, SCF, G-CSF, IL-1beta, and IL-3 seems to be adequate for hematopoietic progenitor/stem cell expansion from cryopreserved cord blood cells.

in vitro Expansion and Differentiation of Cord Blood Hemotopoietic Cells / 中国肿瘤生物治疗杂志

Objective: To explore the role of human hematopoietic growth factors (HGFs) in proliferation and differentiation of cord blood CD34+ cells. Methods: Human cord blood mononuclear cells (MNC) were cultured with different combinations of HGFs (including rhIL-1,rhIL-3, rhIL-6, rhG-CSF, rhGM-CSF and rhSCF). The expansion folds of MNC, the changes of cellular surface markers by FACS, and CFU-GM of hematopoietic cells were observed. Results: The total nucleated cells expanded by 44 folds after cultured with 6 cytokines for 20 days. The number of CFU-GM increased by 14 .74 folds after cultured in liquid for 8 days, but decreased heavily after 18 days. The total number of CD34+ cells increased by 127. 79 ~ 196 .40 folds at 6 to 8 day, but decreased to 101.51 folds at 18 day. Conclusion: Human hematopoietic growth factors can increase the expansion of cord blood CD34+ cells and CFU-GM significantly.

Uso clínico de los factores de crecimiento hematopoyéticos / Clinical use of hematopoyetic growth factor

Los factores de crecimiento hematopoyético {FCH) son producto de la excitante y prometedora industria de la biología molecular y la Ingeniería genética. Se hace una revisión de la farmacología del Factor Estimulador de Colonias de Granulocitos y del Factor Estimulador de Colonias de Granulocitos-Macrófagos, como también de su uso clínico en neutropenia aguda post-quimioterapia mielotóxica anticancerosa, trasplante de médula ósea, leucemia aguda, síndromes mielodisplásicos, anemia aplástica, síndrome de inmunodeficiencia adquirida y neutropenia crónica.

Hematopoietic growth factors are one of the products of the exciting and promising molecular biology and genetic engineering industries. Two of these factors are the recombinant human - granulocyte colony-stimulating factor and the recombinant human-granulocyte-macrophage colony-stimulating factor; a review Is presented on their pharmacology and clinical uses in acute neutropenia after myelotoxic anticancer therapy, bone marrow transplantion, acute leukemia, myelodyplastic syndromes, aplastic anemia, AIDS and chronic neutropenia

Effect of angelica polysaccharide on bone marrow macrophage and its relationship to hematopoietic regulation / 中草药

Objective To investigate the effect of angelica polysaccharide (APS) on bioactivity of bone marrow macrophage (BMM?) and its relationship to hematopoietic regulation, for clarifying the hematonic mechanism of Angelica sinensis. Methods The techniques of hematopoietic progenitor cell culture and BMM? culture in vitro, biological assay of hematopoietic growth factor (HGF) in culture media of BMM?, immunocytochemistry, and nucleic acid in situ hybridization were used. Results The culture supernatant of BMM? induced by APS can enhance the CFU-Mix, CFU-E, CFU-GM; the expression of erythropoietin (EPO), GM-CSF, IL-3, IL-6 protein in BMM? induced by APS was much stronger than that in the control group at different levels; the expression of EPO mRNA and GM-CSF mRNA in BMM? induced by APS were intensified. Conclusion APS may directly and/or indirectly stimulate the BMM? in hematopoietic inductive microenvironment to accelerate the synthesis and secretion of hematopoietic regulation factors on the basis of gene and protein level, such as EPO, GM-CSF, IL-3, IL-6, which in turn to promote the proliferation and differentiation of CFU-Mix, CFU-E, and CFU-GM.