小鼠腹腔巨噬细胞对红系祖细胞增殖的影响

应用造血祖细胞体外培养技术,观察了小鼠腹腔巨噬细胞对早期红系祖细胞(BFU-E)及晚期红系祖细胞(CFU-E)增殖的影响。结果表明,小鼠腹腔贴壁巨噬细胞对CFU-E表现为刺激作用;对BFU-E增殖依培养体系条件不同而表现为刺激和抑制双重作用。巨噬细胞培养上清液也表现相似的作用。提示巨噬细胞具有促红细胞生成素(EPO)和爆式集落促进活性(BPA)样活性。     

不同部位的巨噬细胞培养上清液对小鼠红系血细胞发生的影响

目的:探讨腹腔、肺泡和肝脏的巨噬细胞培养上清液对小鼠早期红系造血祖细胞[用红系爆式集落形成单位(BFU-E)表示]和晚期红系造血祖细胞[用晚期红系集落形成单位(CFU-E)表示]发生的影响.方法:采用造血祖细胞体外培养技术观察3种不同部位的巨噬细胞培养上清液对BFU-E和CFU-E集落数的影响.结果与结论:3种培养上清液均能促进BFU-E和CFU-E的分化与增殖,而肺泡巨噬细胞培养上清液的上述作用更明显.     

人骨髓红系、混合系造血祖细胞无EPO试剂体外培养

红系造血祖细胞(CFU-E、BFU-E)及混合造血祖细胞(CFU-Mix)培养技术对于某些血液病的发病机理的研究、造血细胞分化、增殖及调控方面的研究均有理论上和实践上的意义。红系及混合系祖细胞体外培养的集落形成依赖于红细胞生成素(EPO)的存在,在我     

血液病患者骨髓红系祖细胞体外培养测定

本文采用甲基纤维素培养技术观察了50例血液病患者(男24例,女26例,年龄19～64岁)的骨髓红系祖细胞CFU-E、BFU-E。结果表明,慢性再生障碍性贫血患者骨髓BFU-E和CFU-E集落都减少甚至为重,说明再障是比BFU-E、CFU-E和CFU-D更早阶段的造血干细胞异常所引起的;测定4例急性白血病患者骨髓,其中急淋1例、急粒3例,BFU-E集     

儿童再生障碍性贫血骨髓祖细胞的测定和意义

目的通过对再生障碍性贫血(AA)患儿红系(BFU-E、CFU-E)、粒单核系(CFU-GM)及巨核系(CFU-Meg)祖细胞培养,了解骨髓祖细胞增殖情况.方法抽取47例AA患儿骨髓分别作四类祖细胞培养,在7、14 d测定其集落数.结果 47例AA患儿中,四类集落数均值与对照组比较均有显著下降(P<0.05,P<0.01),且四类集落数均值在AA三种类型中,基本上随病情加重均值逐步下降,其中四类集落数低于对照组下限的患儿分别为BFU-E 36.17%、CFU-E 85.11%、CFU-GM 74.47%、CFU-Meg 91.49%.BFU-E、CFU-GM和CFU-Meg随着患儿疾病的加重,低于对照组下限的患儿百分比同步上升,且三者的表现与外周血三系表现一致,而CFU-E却未见上述现象.结论在部分AA患儿中存在着四类祖细胞培养集落数的下降,且基本上与患儿病情成正比;BFU-E的变化较CFU-E更能代表患儿骨髓红系祖细胞的变化.     

重组人白细胞介素-1β对人红系、粒系造血祖细胞集落形成的影响

为探讨重组人白细胞介素-1β(rhIL-1β)对人红系、粒系造血祖细胞的作用,采用96孔板微量培养法在体外进行培养。在培养中加入不同浓度的rhIL-β1(0～5ug/mL)后,观察BFU-E、CFU-E及CFU-GM的集落形成。结果加入不同浓度的IL-1β后,BFU-E、CFU-E的集落形成均受到抑制,且呈剂量依赖性,在IL-1β剂量为0.1和0.5μg/mL时,抑制作用更明显(JP<0.005,0.05),而IL-1β对CFU-GM的集落形成无明显影响。结果表明,IL-1β对红系造血祖细胞(BFU-E和CFU-E)影响的研究有助于解释某些慢性疾病性贫血(如类风湿性关节炎伴贫血等)的发生机制,提示IL-1β单克隆抗体有可能用于治疗慢性疾病性贫血。     

红系造血祖细胞BFU-E、CFU-E体外培养观察

造血祖细胞培养测定是研究造血干细胞增殖情况的重要方法。BFU-E 和CFU-E 是只能向红系分化的红系造血祖细胞的不同亚群,体外培养时需要添加外源性红细胞生成素(EPO)。1971年Stephenson 以血浆凝块为支持物,首次培养成功人的骨髓红系祖细胞。我们采用甲基纤维素为支持物,培养了人的骨髓红系造血祖细胞BFU-E 和CFU-E,并作了定量测定。光学和透射电子显微镜对BFU-E 和CFU-E 的形态进行了观察,此项工作目前国内尚未见报道。     

rhIL-6,rhGM-CSF和rhEPO对造血干细胞的作用

据《中华内科杂志》1994年33卷第5期报道 北京医科大学第三医院血液科,用三种细胞因子对人正常造血干细胞进行了体外观察。 结果表明:重组人白细胞介素(rhIL-6)和重组人-单细胞集落刺激因子(rhGM-CSF)与正常人造血干细胞培养1周后,IL-6组细胞数增至4.3±0.6倍;CM-CSF组细胞数增至9.4±0.9倍,IL-6 CM-CSF组细胞数增至13.7±1.0倍,明显高于对照组(P<0.01)。CFU-E集落分析结果:IL-6单独应用未见CFU-E集落形成,仅有CFU-GM集落形成;IL-6 EPO(促红细胞生成素)组则CFU-E集落数明显高于EPO组(P<0.01)。CFU-Mix     

冬虫夏草对小鼠红系造血机能的影响

用微量甲基纤维素培养方法探讨冬虫夏草结晶制剂(CS-Cr)对小鼠骨髓红系祖细胞(CFU-E和BFU-E)增殖的影响。实验结果表明,CS-Cr明显提高骨髓CFU-E和BFU-E产率、自杀率,并能对抗三尖杉酯碱对造血机能的损害。采用液体培养技术发现CS-Cr促进骨髓成纤维祖细胞(CFU-F)增殖,其产率变化与CFU-E和BFU-E产率变化一致。     

当归多糖对人红系造血祖细胞增殖的影响及其机理研究

采用造血祖细胞体外培养、造血生长因子生物活性检测等实验血液学技术，研究当归多糖（AP）对人红系造血祖细胞（BFU-E，CFU-E）增殖调控的影响及其机理，结果表明，AP能显著促进BFU-E，CFU-E的集落形成/经AP诱导制备的脾细胞，胸腺细胞和骨髓基质细胞培养上清液能提高BFU-E的集落产率；AP能协同IL-3和红细胞生长素（Epo)促进BFU-E生长，研究提示，AP有可能直接刺激BFU-E，CFU-ZE增殖。亦可能通过促进机体产生红系造敌国生长因子和/或协同这些因子的作用等途径。进而促进机体的红系血发生。     

The effects of total saponins of panax ginseng on hematopoietic progenitor cells in healthy humans and aplastic anemia patients

Ginseng is said to have beneficial effects on anemia. The proliferation effects of total saponins of Panax ginseng (TSPG) on hematopoietic progenitor cell in healthy individuals and 29 patients with aplastic anemia (AA) were observed through bone marrow cultures of burst forming unit-erythroid (BFU-E), colony forming unit-erythroid (CFU-E) and colony forming unit-granulocyte/macrophage (CFU-GM) in vitro compared with methyltestosterone (MT). The results suggest TSPG might prompt the proliferation of normal progenitor cells at a concentration of 20 μg/ml. The numbers of BFU-E,CFU-E and CFU-GM increased by 37.8±2.9%, 31.4±2.9% and 33.3±4.0% respectively overthe controls; furthermore TSPG was still useful to BFU-E, CFU-E growth without Epo in vitro, although the colony numbers were much lower. Otherwise MT was useless to CFU-GM. Of the 29 patients with AA, 14 who responded to MT showed sensitivity to TSPG in marrow culture (the rising rate of colony formation exceeded 30%), but immune-mediated AA (patient’s peripheral blood mononucleated cell suppressed normal hematopoiesis) and stem cell decreased AA (few of colonies were formed) showed almost no expression for TSPG activity because of the immunological suppression system and the absence of progenitors.     

The in vitro growth of erythroid colonies from dog bone marrow

Enriched methyl cellulose media together with either human urinary erythropoietin or serum collected from phlebotomized dogs exposed to hypoxia was used in the study of the erythroid colony forming (CFU-E) capacity of dog marrow. The dog serum erythropoietin was found to be more efficient in stimulating CFU-E than comparable concentrations of human urinary erythropoietin. Numbers of CFU-E were directly related to the culture concentration of the stimulating serum and to the number of cells per plate. Sheep plasma erythropoietin was also found to be effective in stimulating CFU-E growth. The system described is chemically better defined and produced more consistent results than has been reported for the plasma clot method.     

Clinical Observation on Treatment of Chronic Aplastic Anemia by Shengxuening (生血宁) and Cyclosporin A

Objective: To explore the therapy to further elevate the efficacy of the treatment of chronic aplastic anemia (CAA). Methods: Forty-five patients with CCA were assigned into two groups, the 26 paporin A (CsA), and the 19 patients in the control group were treated with androgen alone, with the therapeutic course lasting for over 3 months. Changes of peripheral blood picture, and the colony productivity of burst forming unit-erythroid (BFU-E), colony forming unit-erythroid (CFU-E) and colony forming unit-granulocyte macrophage (CFU-GM) in bone marrow were observed before and after 3 months treatment. The amount of erythrocyte and platelet infusion, frequency of infection, condition of hemorrhage and relevant death were also observed. The follow-up study was conducted for over half a year. Results: The total effective rate in the treated group was 84.6%, which was significantly higher than that in the control group (52.6%, P＜0.05).Levels of hemoglobin, reticulocyte, neutrophil and platelet increased after treatment in the treated group, as compared with those before treatment, with significant difference ( P＜0.05), and the colony productivity of BFU-E, CFU-E and CFU-GM in bone marrow also got significantly increased ( P＜0.01 ), and showed significant difference from those in the control group (P＜0.05). Conclusion: Shengxuening-assisting CsA therapy is an effective measure for treatment of CAA.     

Clinical observation on treatment of chronic aplastic anemia by Shengxuening and cyclosporin a

Objective: To explore the therapy to further elevate the efficacy of the treatment of chronic aplastic anemia (CAA).Methods: Forty-five patients with CCA were assigned into two groups, the 26 patients in the treated group were treated by Shengxuening (a Chinese herbal preparation) and cyclosporin A (CsA), and the 19 patients in the control group were treated with androgen alone, with the therapeutic course lasting for over 3 months. Changes of peripheral blood picture, and the colony productivity of burst forming unit-erythroid (BFU-E), colony forming unit-erythroid (CFU-E) and colony forming unit-granulocyte macrophage (CFU-GM) in bone marrow were observed before and after 3 months treatment. The amount of erythrocyte and platelet infusion, frequency of infection, condition of hemorrhage and relevant death were also observed. The follow-up study was conducted for over half a year.Results: The total effective rate in the treated group was 84.6%, which was significantly higher than that in the control group (52.6 %,P< 0.05). Levels of hemoglobin, reticulocyte, neutrophil and platelet increased after treatment in the treated group, as compared with those before treatment, with significant difference (P< 0.05), and the colony productivity of BFU-E, CFU-E and CFU-GM in bone marrow also got significantly increased (P< 0.01), and showed significant difference from those in the control group (P< 0.05).Conclusion: Shengxuening-assisting CsA therapy is an effective measure for treatment of CAA.     

Effects of Weiganfi on the Hemopoietic Function of the Myelosuppressed Anemic Mice

To study the effect of Weiganli （维肝力） on bone marrow hemopoiesis. Methods： The effects of Weiganli on the peripheral blood picture and the number of bone marrow nucleated cells （BMCs） were observed in myelosuppressed anemic model mice, and the effects of Weiganli on the growth of colony forming unit-granulocyte macrophage （CFU-GM）, colony forming unit-erythroid （CFU-E）, burst forming unit-erythroid （BFU-E）, colony forming unit-megkaryocyte （CFU-Meg） were investigated by in vitro cell culture technique. The hemopoietic stem cells （HSCs, c-kit＋） in bone marrow were double stained with fluorescent antibody PE-C-Kit and FITC-CD45, and the HSCs （c-kit＋） were counted by flow cytometer with CD45/SSC （side scatter） gating. Results： Peripheral blood cell counts and the number of BMCs were significantly improved after Weiganli administration; and bone marrow hemopoietic stem/progenitor cells were significantly increased. Conclusion： Weiganli can effectively promote the recovery of hemopoietic function in the mvelosuooressed anemic mice.     

Ⅱ型登革病毒对骨髓红系造血细胞的影响

用体外造血祖细胞培养技术证明Ⅱ型登革病毒对红系造血祖细胞(CFUE)也有生长抑制作用,CFU-E的抑制效应与病毒的浓度呈正相关。说明登革热病人虽然无贫血表现,但骨髓红系造血已受到病毒的影响。     

小鼠骨髓巨核祖细胞体外微量培养

本文首次报道采用微量甲基纤维素体外培养法,培养出小鼠骨髓巨核祖细胞。1u/ml促红细胞生成素、5％植物血凝素条件培养液及2.5％美州商陆素条件培养液对提高巨核细胞产率最为适宜。     

干扰素γ介导噬血细胞综合征血细胞减少的机制研究

目的探讨噬血细胞综合征患者与正常人骨髓CD34＋细胞质量的差异情况,研究干扰素γ对骨髓分化的影响和特点,并观察提高集落刺激因子对干扰素γ抑制骨髓作用的对抗情况。方法采用免疫磁珠法分离纯化骨髓CD34＋造血干细胞。在体外扩增培养中,以干细胞因子（stem cell factor,SCF）和白介素-3（interleukin-3,IL-3）、粒细胞-巨噬细胞集落刺激因子（granulocyte macrophage-colony stimulating factor,GM-CSF）、粒细胞集落刺激因子（granulocyte-colony stimulating factor,G-CSF）、促红细胞生成素（erythropoietin,EPO）集落刺激因子组合对骨髓干细胞诱导分化,以集落形成单位（colony forming units,CFU）的形成数来评价骨髓分化情况,在体系中加入不同浓度梯度的干扰素γ,观察骨髓集落形成情况。在干扰素γ抑制骨髓的最低浓度下增加集落刺激因子浓度,观察骨髓生长情况。结果患者骨髓CD34＋细胞质量与正常人相比差异无统计学意义（P〉0.05）;不同浓度的干扰素γ对骨髓的抑制程度不同,当干扰素γ浓度提高到10 ng/mL以上时,实验组的集落数目相比于未添加干扰素γ的对照组,差异有统计学意义（P〈0.05）;增加培养体系中集落刺激因子浓度可对抗因干扰素γ使骨髓出现的抑制作用情况。结论噬血细胞综合征患者与正常人骨髓质量不存在差异,干扰素γ可以对骨髓生长起抑制作用,提高集落刺激因子浓度可以对抗干扰素γ产生的抑制作用。     

Myeloid and erythroid hemopoiesis supported by human bone marrow fibroblasts in vitro.

The passages 0 and 3 purified human bone marrow fibroblast layers (FLs) were established by long-term liquid cultures. After 12-day coculture of stromal. cell-depleted marrow cell suspensions with passage 0 FLs, 68.33 +/- 4.04% of the hemopoietic colonies adhered to FLs was myeloid in nature and the other 31.67 +/- 4.04% was erythroid. There were still CFU-E (colony forming unit-erythroid), BFU-E (burst forming unit- erythroid), and CFU-GM (colony forming unit-granulocyte/macrophage) among the nonadherent cells. The media conditioned by passage 0 (F0-CM) and passage 3(F3-CM) FLs stimulated the growth of myeloid and erythroid (BFU-E) colonies. From these data, it is concluded that both FLs and the media conditioned by fibroblasts can stimulate myeloid and erythroid hemopoiesis.