Radiosensitivity of colony-forming units of dog bone marrow in agar cultures

The radiosensitivity of colony-forming units in dog bone marrow was determined by a modified method of cloning hematopoietic cells in semisolid agar gel in diffusion chambers in vivo. The dose of radiation whose action was followed by preservation of 37% of the objects (D_o) was 144±14.8 rad (n=0.8) for committed precursor cells of granulocytopoiesis (CFUc) and 468±35.8 rad (n=0.9) for precursor cells forming stellate colonies of fibroblast-like cells (CFUf). It is concluded that the CFUf belong to the class of stromal precursors of hematopoietic organs. This system is suitable for the simultaneous study of hematopoietic and stromal precursor cells in dogs.Laboratory of Bone Marrow Culture and Transplantation, Central Institute of Hematology and Blood Transfusion, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR N. A. Fedorov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 87, No. 4, pp. 340–342, April, 1979.     

小鼠同种异基因骨髓腔内骨髓移植促进早期造血功能重建

目的探讨同种异基因骨髓腔内骨髓移植(IBM-BMT)对小鼠早期造血功能重建的影响。方法将BALB/c小鼠骨髓单个核细胞(BMNCs)分别用胫骨骨髓腔内注射(IBMI)和尾静脉注射(IV)两种方法移植入经致死量60Coγ射线辐照后的60只C57BL/6小鼠。受鼠随机分为3组:骨髓腔内注射高和低剂量组(IBM1和IBM2组)、尾静脉注射组(IV组),每组20只。在骨髓移植后1、3、6和9d分别计数各组受鼠胫骨骨髓腔内有核细胞总数,并用流式细胞术检测供体植入水平(供体来源有核细胞总数、供体来源髓系细胞数)。结果于移植后6d,IBM1组和IBM2组注射侧胫骨骨髓腔内有核细胞总数、供体来源有核细胞总数、供体来源髓系细胞总数均明显高于IV组(P<0.05或P<0.01?。结论IBM-BMT较IV-BMT更能促进同种异基因骨髓移植后的早期造血功能重建。     

人参总皂甙对骨髓巨噬细胞的影响及与造血调控的关系

目的：研究人参总皂甙(TSPG)对骨髓巨噬细胞(BMMφ)生物学活性的影响及其与造血调控关系。方法：采用骨髓造血祖细胞体外培养,造血生长因子生物活性检测,免疫细胞化学,核酸探针原位杂交等技术。结果：经TSPG诱导制备的BMMφ的培养上清可提高髓系造血祖细胞的集落产率;经TSPG诱导后,BMMφ表达EPO,IL-3,IL-6,GM-CSF的蛋白水平有不同程度提高;EPO mRNA,GM-CSF mRNA的表达水平和强度明显提高。结论：TBPG可以刺激造血诱导微环境中的巨噬细胞,从基因水平和蛋白水平2级层次上促进造血调控因子的合成和分泌,进而促进CFU-Mix,CFU-E,CFU-GM的增殖分化。     

Effect of platelet growth factors on proliferation of guinea pig bone marrow and splenic stromal precursor cells and proliferation of cultural descendants from bone marrow precursor cells

Elimination of platelets from guinea pig splenocyte suspension (laking megakaryocytes) with EDTA considerable reduces the efficiency cloning of splenic stromal precursor cells. It means that platelet-derived growth factors are necessary for stromal precursor cells from different organs (bone marrow, thymus, and spleen). The dependence on the platelet growth factors can vary within a wide range in descendants from cultured bone marrow precursor cells (passaged bone marrow fibroblasts at different stages of differentiation.     

血液血管细胞生成素对胎儿骨髓造血和内皮干细胞作用的研究

目的研究人血液血管细胞生成素(hemangiopoietin,HAPO)对胎儿骨髓细胞的作用,探讨其生物学特性。方法采用细胞液体培养、半固体培养、MTT方法、免疫荧光标记流式细胞仪测定、免疫组化、显微镜观察照相等方法。结果在液体培养3周的胎儿骨髓单个核细胞中,HAPO组中出现了大量小而圆的早期造血细胞,其中CD34+细胞含量比对照组高20%,对照组CD34+细胞为1.25×105个,而HAPO组CD34+细胞为3.93×106个。取胎儿骨髓悬浮造血细胞进行半固体培养,对照组不能形成CFU-GEMM,而HAPO组形成CFU-GEMM数达到(11.0±2.6)个;HAPO也协同SCF、IL-3、GM-SCF等生长因子促进集落形成,CFU总数是对照组2.6倍,CFU-GEMM数HAPO组是对照组2.1倍。MTT方法发现,HAPO对胎儿骨髓基质细胞也有促增殖作用,HAPO可使基质细胞增长21%;液体培养的胎儿骨髓基质细胞中,有内皮特异性标志的细胞均增高;在甲基纤维素半固体培养中HAPO使胎儿骨髓内皮细胞的集落数增高,并出现条索状排列的集落,有促进血管形成的趋势。进一步证明HAPO可直接促进CD34+KDR+细胞的增殖。结论HAPO对骨髓造血和血管内皮干细胞均有刺激增殖作用。     

人脐血间充质干细胞移植修复骨髓造血损伤

文题释义：间充质干细胞：是一种多功能干细胞,由胚胎发育早期的中胚层发展而来,存在于人的各种组织、器官中,如骨、软骨、脂肪、外周血和肌肉等。骨髓组织中的间充质干细胞最多,但其在骨髓细胞中的比例仍很低,只有0.01%-1.00%,且年龄越大其含量越少,骨髓中的间充质干细胞分化能力也呈下降趋势。与骨髓相比,脐血中所含的间充质干细胞更加原始,因而有更强的增殖、分化能力。相较于骨髓间充质干细胞而言,人脐血间充质干细胞具有来源广泛、取材方便、无伦理方面的限制,使得人脐血间充质干细胞成为再生医学中的另一重要来源。骨髓造血损伤动物模型：建立骨髓造血损伤动物模型的方法有很多,主要分为物理方法、化学方法及物理化学方法等。物理方法包括各种射线,如X射线等,化学方法主要指以环磷酰胺为代表的烷化剂类化疗药物,而物理化学方法也叫混合性方法,联合应用放射线和化疗药物建立动物模型。  摘要背景：大多数研究间充质干细胞体外培养对造血干细胞的增殖作用和骨髓间充质干细胞移植可降低辐照引起的造血细胞死亡,增加骨髓细胞存活,修复造血功能,而少有研究人脐血间充质干细胞移植对骨髓造血损伤的修复。目的：探讨人脐血间充质干细胞对骨髓造血微环境的修复情况。方法：选用雄性BALB/c小鼠随机分为3组,实验组和对照组小鼠进行总剂量为6 Gy的X射线全身照射,建立骨髓造血损伤模型,正常组为未经处理的正常小鼠。实验组小鼠照射当天经尾静脉输入CM-DiL标记的人脐血间充质干细胞5×10⁶/只(0.2 mL),对照组和正常组经尾静脉输入生理盐水0.2 mL,移植后第1,5,7,14,21天观察外周血血象恢复情况和骨髓造血微环境修复情况。结果与结论：①外周血常规：移植后第1,5,7天,实验组和对照组小鼠与正常组小鼠比较,白细胞、血小板、红细胞计数及血红蛋白浓度进行性下降,第7天下降最为明显,移植后第14天三系较前有所恢复,移植后第21天基本恢复正常,与实验组相比,对照组三系下降更为明显,移植后第14天实验组较对照组恢复快;②骨髓涂片情况：移植后第1,5,7,14天实验组及对照组小鼠骨髓出现造血功能抑制,以第7天最为明显,移植后第14天骨髓增生较前有所恢复,实验组优于对照组;移植后第21天实验组及对照组小鼠骨髓造血功能恢复,与正常组相比无差异;③骨髓病理切片情况：移植后第1,5,7,14天实验组及对照组小鼠骨髓出现造血功能抑制;移植后第14天实验组及对照组小鼠的骨髓造血功能较前开始恢复,实验组小鼠的骨髓增生情况优于对照组小鼠, 移植后第21天实验组及对照组小鼠骨髓增生情况与正常组比较无差异;④结果表明,人脐血间充质干细胞对骨髓造血功能恢复均有明显促进作用。ORCID: 0000-0002-7547-9664(高坤莉) 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

Comparative analysis of the action of unpurified mouse erythropoietin and human recombinant erythropoietin on erythroid and granulocytic-macrophagal precursor cells in semisolid mouse bone marrow cultures

Hematology Research Center, Ministry of Health of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. I. Vorob'ev.) Translated from Byulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 112, No. 8, pp. 176–179, August, 1991.     

衰老模型骨髓基质细胞抑制造血细胞增殖分化

目的研究衰老骨髓基质细胞对骨髓造血细胞增殖分化能力的影响,为阐述机体造血微环境衰老对造血干/祖细胞增殖的影响提供实验依据。方法全骨髓贴壁法体外培养大鼠骨髓基质细胞,分为对照组和衰老组。衰老组:常规培养基内加入30 mg/m L D-半乳糖作用48 h。CCK-8法测定BMSCs增殖;流式细胞术分析细胞周期;β-半乳糖苷酶(SA-β-Gal)染色观察衰老BMSCs百分率;Western blot检测P16、P21和P53蛋白表达。骨髓造血细胞与BMSCs共培养,集落计数检测髓系多向性造血祖细胞(CFU-Mix)增殖分化。ELISA检测BMSCs培养上清液中IL-1β、GM-CSF和SCF含量;DCFH-DA流式荧光检测BMSC活性氧簇(ROS)水平;酶学法检测BMSCs内过氧化物丙二醛(MDA)含量和总超氧化物歧化酶(SOD)活性。结果与对照组相比,D-半乳糖诱导BMSCs衰老,细胞阻滞于G0/G1期(P0.01),增殖能力显著下降,SA-β-Gal染色阳性率升高(P0.01);衰老相关蛋白P16、P21和P53表达明显上调(P0.01)。与衰老BMSCs共培养的骨髓造血细胞增殖分化能力减弱。衰老BMSCs内ROS、MDA氧化损伤指标上升,SOD抗氧化指标下降(P0.01);BMSCs培养上清液IL-1β、GM-CSF和SCF含量明显下降(P0.01)。结论衰老骨髓基质细胞抑制造血细胞增殖、分化能力,其机制可能与骨髓基质细胞氧化损伤,分泌活性因子改变有关。     

Ghrelin抑制小鼠骨髓基质细胞成脂分化

目的观察胃促生长素ghrelin对原代培养小鼠骨髓基质细胞成脂分化的影响。方法体外培养小鼠骨髓基质细胞,MDI诱导剂诱导细胞成脂分化,显微镜观察细胞形态变化;油红O染色法检测细胞甘油三酯含量,化学比色法测定碱性磷酸酶(ALP)活性;MTT法检测细胞克隆化扩增活动,RT-PCR检测过氧化物酶体增殖物激活受体γ2(PPARγ2)和丝氨酸蛋白酶脂肪因子adipsinmRNA表达,Westernblot检测PPARγ2蛋白表达。结果Ghrelin能够剂量依赖性(10-9、10-8、10-7mol/L)地抑制骨髓基质细胞成脂分化,升高ALP活性(P<0·05或P<0·01)。Gh-relin能够降低PPARγ2和adipsinmRNA以及PPARγ2蛋白表达(P<0·05)。Ghrelin对细胞成脂分化早期的克隆化扩增没有显著影响。结论Ghrelin能够显著抑制体外培养小鼠骨髓基质细胞成脂分化,该作用可能与抑制PPARγ2表达有关。     

Damage to cell DNA in the bone marrow and testes of mice with experimental trichinosis

Metabolites ofTrichinella spiralis produced genotoxic and cytotoxic effects on somatic and generative cells of the host organism. They increased the number of single-chain breaks, alkaline-labile sites in nuclear DNA, and count of apoptotic cells in the bone marrow and testes of infected mice. These effects depended on the stage of parasite development in the host organism and became more pronounced with increasing invasion intensity. Translated fromByulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 138, No. 9, pp. 320–323, September, 2004     

Damage to cell DNA in the bone marrow and testes of mice with experimental trichinosis

Metabolites of Trichinella spiralis produced genotoxic and cytotoxic effects on somatic and generative cells of the host organism. They increased the number of single-chain breaks, alkaline-labile sites in nuclear DNA, and count of apoptotic cells in the bone marrow and testes of infected mice. These effects depended on the stage of parasite development in the host organism and became more pronounced with increasing invasion intensity.Translated from Byulleten Eksperimentalnoi Biologii i Meditsiny, Vol. 138, No. 9, pp. 320–323, September, 2004     

Clonogenic methods in vitro for the enumeration of granulocyte-macrophage progenitor cells (CFU-GM) in human bone marrow and mouse bone marrow and spleen

Summary Protocols and techniques are described for the in vitro enumeration of granulocyte and macrophage progenitors found in human bone marrow and mouse bone marrow and spleen. Study of the colony forming unitgranulocyte-macrophage (CFU-GM) in human bone marrow is usually first accomplished by density separation of whole bone marrow buffy coats through Ficoll-Paque into low- and high-density fractions. After collection of the low-density fraction (containing most or all of the CFU-GM), cells are washed and suspended, at a known cell concentration, in a mixture of culture medium, fetal bovine serum (or serum-supplements), agar or agarose, with a source of colony stimulating factor(s). Cultures are allowed to solidify and are then placed in a humidified 37°C incubator at 5% CO₂ in lowered (5%) O₂ tension for 7 to 14 d. Colonies (>40 cells/ aggregate) and clusters (3 to 40 cells/ aggregate) are then scored. Murine CFU-GM are cultured and characterized in a similar manner except tissues are separated into a single cell suspension without a density separation, and the culture time is reduced to 5 to 7 d. Colonies and clusters are scored as previously described. Purification of CFU-GM can be performed and these cells cultured as above.