体外半固体琼脂前红系祖细胞微量培养实验观察

采用半固体琼脂微量培养方法培养人骨髓红系造血祖细胞集落获得成功。培养体系中不需条件培养基和细胞生长刺激剂。显微镜下红、粒系集落区别明显,细胞形态清晰,方法简易,减少了所需细胞数量和节省了试剂,特别可节省促红细胞生成素。     

国产红细胞生成素对人体胎肝细胞形成红系集落的刺激作用

国产红细胞生成素(EPO)刺激妊娠4～5个月的胎肝红系祖细胞(BFU-E、CFU-E)形成集落,CFU-E集落的峰值出现于培养后48小时,BFU-E在7～10天借助于血红蛋白的特征可直接观察。CFU-E的产率与种入的细胞数呈线性关系,并与EPO浓度密切相关,100mu/ml培养物时集落计数最高。     

关于正常人及献血员循环红系祖细胞测定的初步观察

应用体外培养方法外加商品红细胞生成素(EPO)或再生障碍性贫血病人血清(AS)对5名正常人和6名5 年以上献血员的外周血红系祖细胞进行检测,证明正常人循环中存在两类红系祖细胞,献血员的后红系祖细胞含量正常,而前红系祖细胞明显低于正常。加 EPO与加AS的培养结果比较,差别无显著性。     

重组人类促红细胞生成素对尿毒症透析患者骨髓红系集落形成单位的影响(附11例病例分析)

分析11例重组人类促红细胞生成素对尿毒症透析患者骨髓有核红细胞和红系集落形成单位(CFU-E)的影响。结果表明,用药前CFU-E比正常对照明显下降(p<0.01),用药40～65d后CFU-E较用药前明显升高(p<0.05),同时骨髓有核红细胞数较用药前亦有所增多。认为重组人类促红细胞生成素能升高尿毒症透析者骨髓CFU-E水平,促进其增殖分化,从而改善肾性贫血。     

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。     

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     

分子伴侣在红细胞生成过程中的作用

由髓系造血干/祖细胞开始的红细胞造血经历了大约3个阶段,首先是定向分化为红系爆式集落形成单位(BFU-E),然后分化为红系祖细胞集落形成单位(CFU-E),最后逐渐成熟,成为形态上可辨认的幼稚红细胞到去核红细胞.该过程受到多种因子调控并伴有复杂的胞内信号转导.     

非EPO依赖性红系祖细胞培养与真性红细胞增多症

真性红细胞增多症(PV)是一种以红细胞克隆性增生为主的慢性骨髓增生性疾病,其病因和发病机制至今尚不明确.文献报道大多数PV患者存在非促红细胞生成素(EPO)依赖性红系祖细胞生长,并推测它可能与PV的发生有一定关系.我们对21例临床诊断为PV的患者进行了EPO依赖性和非EPO依赖性红系爆式集落形成单位(BFU-E)培养,探讨其与PV的关系.     

血管紧张素Ⅱ对造血祖细胞优势扩增的影响

目的 探讨血管紧张素Ⅱ(AngⅡ)及其受体拮抗剂氯沙坦对不同系别造血祖细胞分化的影响. 方法 悬浮培养的单个核细胞分为5组:促红细胞生成素(EP0)组(Ⅰ组),EFO AngⅡ组(Ⅱ组),EPO AngⅡ 氯沙坦组(Ⅲ组),AngⅡ组(Ⅳ组)及对照组(Ⅴ组).巢式逆转录聚合酶链反应(nested-RT-PCR)检测血管紧张素Ⅱ1型受体(AT1R)基因的mRNA表达.计数红系爆式集落形成单位(BFU-E)和粒一单系集落形成单位(CFU-GM). 结果 ①EPO组及EPO AngⅡ组于第6,9天可检测到AT1R mRNA表达;在相同时间点EPO AngⅡ组较EPO组AT1R的mRNA表达量增多.②EPO AngⅡ组较其余各组红系集落明显增多,粒系集落明显减少.AngⅡ组及对照组均未检测到红系集落. 结论 AngⅡ在EPO存在下与AT1受体结合促使红系祖细胞向红系优势分化.氯沙坦通过抑制AngⅡ与AT1受体结合,抑制红系分化.     

肾上腺素影响小鼠粒巨噬系祖细胞生长的实验研究

目的了解肾上腺素(Adrenaline,Adr)对小鼠粒巨噬系祖细胞的影响.方法在体外半固体培养的粒-巨噬系集落形成单位(Colony Form Unit-Granulocyte and Macrophage,CFU-GM)中加入不同浓度的Adr后,计数集落数,并与阴性对照组比较.结果Adr浓度在10-6～10-10mol/ml范围内,CFU-GM集落数与对照组相比,P＞0.05;Adr浓度为10-5mol/ml时,P＜0.05;Adr浓度为10-4mol/ml时,培养体系中没有CFU-CM形成.结论高浓度Adr可抑制CFU-GM在体外的生长,儿茶酚胺类物质能参与血发生的调节.     

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 4 7例AA患儿骨髓分别作四类祖细胞培养 ,在 7、14d测定其集落数。结果 4 7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 .0 5 ,P <0 .0 1) ,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 .4 7%、CFU Meg 91.4 9 %。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。     

组胺受体激动剂及其拮抗剂对红系祖细胞产率的影响

研究组胺及其拮抗剂对早期和晚期红系祖细胞(BFU-E 和CFU-E)产率的影响。结果表明,一定浓度的组胺和2—噻唑乙胺对CFU-E和BFU-E的产率均产生抑制效应,而4—甲基组胺则无此作用;事先用甲苯苄二胺阻断H_1-受体,能减弱组胺对CFU-E和BFU-E的抑制效应;相同条件下用甲氰咪胍阴断H_2-受体无此效应。提示组胺对红系祖细胞的抑制效应系通过组胺H_1—受体所介导。     

三尖杉酯碱对红系造血机能的影响

用微量甲基纤维素培养方法检测三尖杉酯碱(Harringtonine,Ha)促进小鼠骨髓红系祖细胞CFU-E和BFU-E增殖的作用。结果表明,适当剂量的Ha能增强骨髓红系造血机能;如剂量过大反而起抑制作用。双盲实验也表明Ha有促进骨髓CFU-E和BFU-E增殖的作用。     

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.     

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。     

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.     

腹腔巨噬细胞培养上清液对小鼠红系造血调控的研究

本文研究了未刺激和经大肠杆菌内毒素(LPS)刺激的腹腔巨噬细胞培养上清液对小鼠红系造血的调控作用。结果提示:未刺激和经LPS刺激腹腔巨噬细胞培养上清液对早期红系祖细胞(BFU-E)均呈刺激作用,对晚期红系祖细胞(CFU-E)呈双向调控,即低浓度刺激,高浓度抑制。二者比较,LPS刺激腹腔巨噬细胞培养上清液具有更高调控活性。实验还提示,腹腔巨噬细胞培养上清液具有促红细胞生成素(EPO)、BPA(Burst Promoting Activity)样活性     

再生障碍性贫血患者造血祖细胞及免疫机制的研究

目的探讨再生障碍性贫血(再障)的发病机制。方法采用甲基纤维素及胶原半固体培养法对56例再障骨髓CFU-GM、CFU-E、BFU-E及CFU-M K、BFU-M K进行培养;采用APAAP法及双抗体夹心酶联免疫吸附法(EL ISA)对再障患者外周血T细胞亚群、血清sIL-2R进行检测。结果再障患者CFU-GM、CFU-E、BFU-E及CFU-M K、BFU-M K集落数均低于对照组,且严重型再障集落数减少程度与慢性再障相近;再障患者CD 3亚群细胞无变化,CD 4亚群细胞减低,CD 8亚群细胞增高,CD 4/CD 8降低,重型再障患者血清sIL-2R中水平高于正常对照组与慢性再障。结论骨髓造血祖细胞数量减少是再障发病的重要因素,其减少程度与再障的病情有关;细胞免疫功能异常及造血负调控因子可能在再障发病中起一定的作用。     

Bovine granulocyte/macrophage and erythroid colony culture: characteristics of the colonies and the assay systems.

Bovine bone marrow granulocyte/macrophage colonies were cultured in vitro in methyl cellulose and in plasma clots using bovine endotoxin-stimulated serum as a source of colony stimulating activity. The endotoxin-stimulated serum was four times as potent as the control serum in the methyl cellulose cultures. No significant increase in the number of colony forming units was observed when bovine marrow cells were maintained in suspension cultures for various periods prior to plating in methyl cellulose. The percentage of glass/plastic adherent cells in bovine marrow cells was observed to be 43% +/- 12 (SD). Benzidine positive erythroid colonies appeared in plasma clot cultures on day 4 and disappeared by day 9. No second population of erythroid colonies appeared either as a function of time or as a function of erythropoietin concentration. The optimum erythropoietin concentration for bovine erythroid cultures was found to be 1.0 unit/mL. A significant difference was observed between animals in their marrow capacity to produce erythroid colonies in culture but no significant difference was observed within individual animals over a period of three months.