全反式维甲酸对骨髓纤维细胞的影响

通过纤维细胞集落(CFU-F)的培养,观察了急性早幼粒细胞性白血病(APL)的CFU-F,结果APL 患者较正常骨髓明显低下(4.3±1.63/2×10~5细胞)(n=9);完全缓解后则上升达正常水平(12.4±2.55/2×10~5)。体外试验,维甲酸(RA)加入CFU-F 培养系后,CFU-F 下降为3.11±1.24/2×10~5细胞,明显低于对照组8.5±1.57/2×10~5(P<0.05)。提示RA 对骨髓纤维细胞有抑制作用,此结果可能为RA 治疗骨髓纤维化提供初步依据。     

Clinical significance of human bone marrow stromal cell colonies in acute leukemias

Human bone marrow-derived fibroblastoid colony-forming cells (CFU-F) and adipocyte colonies in 36 patients with acute leukemia were studied, and were serially analysed at different clinical stages. At untreated stage, CFU-F number in acute lymphoblastic leukemia (ALL) was lower than that in acute non-lymphoblastic leukemia (ANL). In ANLs, CFU-F number in M1 was lower than that in M2. Adipocyte colonies were frequently developed at regenerating and relapsing stages, but rarely at untreated and remission stages. The adipocyte colony formation did not correlate with any of CFU-F number, marrow cellularity nor number of leukemic cells, but might be associated with hemopoietic regeneration. The favorable prognosis was associated with normal CFU-F number and with adipocyte colony formation at regenerating or bottom stage. As adipocytes in marrow samples were completely removed before cultures, adipocyte colony was probably originated from preadipocytes. Thus, our results suggest that adipocyte precursor cells increase in regenerating marrow and that they are essential in active hemopoiesis.     

自体骨髓基质细胞回输促进急性白血病病人化疗后骨髓造血恢复的研究

目的通过回输成纤维集落生成单位（CFU—F）促进骨髓实质细胞造血恢复。减轻白血病化疗的副作用，促进骨髓损伤早日恢复。方法选取经明确诊断的AML患者化疗回输组前后行自身对照，化疗回输前及后采骨髓行红系造血祖细胞（BFU-E）、粒、巨系集落生成单位（CFU—GM）培养，观察化疗回输前后的外周血象。结果化疗回输CFU-F后第3周。其BFU—E、CFU-GM数明显高于化疗回输前的水平（P〈0．05）。同时高于单纯化疗组第3周测得的BFU-E和CFU-GM数（P〈0．05）。结论体外扩增造血基质细胞于化疗自体回输方法，促进了病人造血恢复，对预防和减轻化疗药物造成的骨髓损伤，缩短化疗间期，加大化疗药物剂量，具有理论意义和应用价值。     

骨髓增生异常综合征及慢性原因不明性粒细胞减少症患者骨髓CFU—GM和CFU—F的初步观察

利用人类骨髓粒一单核系祖细胞集落(CFU—GM)和成纤维细胞集落(CFU—F)体外检测技术,比较观察32例骨髓增生异常综合征(MDS)和11例慢性原因不明性粒细胞减少症(CIN)患者的造血改变。结果表明,MDS 和 CIN 的造血状态存在显著区别,69%的 MDS 患者 CFU—GM集落显著受抑,集簇、簇/落比值明显升高;其 CFU—F 集落亦存在明显缺陷。而 CIN 患者的上述指标均类似于正常。这提示体外集落检测技术有助于 MDS 和 CIN 的鉴别。此外,CFU—GM 系列培养对 MDS 患者具有一定预后价直。     

Reactive oxygen species produced in mitochondria are involved in age-dependent changes of hematopoietic and mesenchymal progenitor cells in mice. A study with the novel mitochondria-targeted antioxidant SkQ1

Lifelong treatment of mice with the effective mitochondria-targeted antioxidant SkQ1 [10-(6′-plastoquinonyl) decyltriphenylphosphonium] does not affect hematopoietic stem cells (HSC) and more differentiated hematopoietic progenitors but significantly decelerates age-dependent changes in peripheral blood. During the first 13 months, SkQ1 (0.9 or 28.8 nmol/kg day) prevents age-dependent myeloid shift (increase in the proportion of granulocytes and decrease in the proportion of lymphocytes). During the next year of treatment the effect disappears, and the hemogram of 2-year-old treated mice does not differ from the control. The number of mesenchymal stem cells (MSC) in the bone marrow does not change during 2 years of treatment with SkQ1, but the concentration of MSC progeny fibroblast colony-forming units (CFU-F) increases with dose of SkQ1. The concentration of CFU-F after 1 and 2 years treatment with SkQ1 is twice higher than in young mice. Our data indicate that the stromal environment of hematopoietic cells could be the primary target of age-dependent changes mediated by reactive oxygen species produced in mitochondria. The anti-aging effects of SkQ1 described here are in perfect agreement with the inhibitory effects of this antioxidant on aging observed in the other models.     

豚鼠骨髓滋养层对CFU-F的影响及其最佳作用时间的研究

本文研究了受60Gy照射的豚鼠骨髓细胞作滋养层对正常小鼠骨髓细胞造血基质祖细胞(CFU-F)体外培养成集落的影响,优选了滋养层与骨髓细胞的最佳作用时间。种植骨髓细胞当天或次日添加滋养层细胞后继续培养7～10天则CFU-F数最多;种植骨髓细胞当天加滋养层而次日通过换液去掉未贴壁细胞则CFU-F数最低;骨髓细胞培养4天后再加滋养细胞则不起成集落刺激作用;不论在集落数及集落所含细胞数上,加滋养层者都比不加者为多。     

CFU-F from dog marrow: A colony assay and its significance

Summary A colony assay method is described for studying dog fibroblast colony development in marrow cells derived from resected ribs. The assay showed an increased number of fibroblast colony forming units (CFU-F) in cell suspensions prepared from resected ribs compared to cell suspensions prepared from bone marrow aspirates or from peripheral blood. A linear relationship between the number of cells plated and the number of fibroblastoid colonies was demonstrated in each case. The proportion of phagocytic cells was lower in cultures prepared from resected ribs than in those prepared from bone marrow aspirates. Staining for acid phosphatase and with sudan black showed differences between phagocytic cells and non-phagocytic fibroblasts. When left in plastic dishes for 2 hrs, 81% ± 10% of the CFU-F adhered to the plastic dishes. Velocity sedimentation separation showed a modal sedimentation rate of 6.49 mm/h.     

Effects of chemotherapy on bone marrow stroma in mice with acute myelogenous leukemia. Correlation with CFU-C and CFU-D

This study describes changes in bone marrow stroma in murine acute myelogenous leukemia (AML). The AML was induced in C57B1 mice by intravenous (i.v.) transfusion of C4198 myelogenous leukemic cells. In untreated leukemic mice, the colony-forming unit fibroblasts (CFU-F) were severely inhibited. In leukemic mice treated by three chemotherapy protocols of cytosine-arabinoside (Ara-C) and adriamycin there was a 200% increase in the life span as compared to untreated leukemic animals and marked reduction of marrow leukemic load. In these mice the stromal inhibition was temporarily relieved, expressed by peaks of CFU-F2-3 days following each protocol. In between the peaks, CFU-F decreased to subnormal levels, remaining low to the end of the disease. In normal mice administered a similar chemotherapy regimen, there were peaks of CFU-F activation after each protocol and normal levels in between the peaks. Granulocyte/macrophage progenitors (CFU-C) of leukemic-treated and normal-treated mice showed increased levels following each chemotherapy protocol. Whereas CFU-C decreased below normal levels in leukemic mice towards the end of the disease, the level of these progenitors remained high in normal mice receiving Ara-C and adriamycin. Colony-forming units in diffusion chamber (CFU-D) showed mild fluctuations in both leukemic and normal mice receiving three protocols of Ara-C and adriamycin. It is possible that despite treatment, the bone marrow stroma in leukemia becomes irreversibly deficient towards the end of the disease and cannot support the residual normal hematopoiesis.     

骨髓内皮细胞分泌的抑制CFU—F生长的因子

为探讨骨髓微环境中内皮细胞调节造血的可能机制,本实验研究了骨髓内皮细胞无血清条件培养液（ＢＭＥＣ－ＣＭ）及其不同分子量组分对骨髓成纤维祖细胞（ＣＦＵ－Ｆ）生长的作用。在体外培养条件下,收集人和小鼠纯骨髓内皮细胞无血清条件培养液,应用连续超滤技术从其中制备出分子量〉１０ｋＤ,３－１０ｋＤ和〈３ｋＤ三种组分,进行骨髓ＣＦＵ－Ｆ集落形成实验,检测这些条件培养液及其超滤组分的作用。结果显示,人和小鼠骨髓内     

人参蜂王浆对正常和照射豚鼠造血免疫器官CFU-F影响的实验研究

骨髓及脾细胞体外培养中加不同浓度人参蜂王浆后培养8～12天,计数基质祖细胞集落(CFU-F)数。发现实验组形成的CFU-F小型集落明显高于对照组。培养1Gy照射的骨髓细胞,加入3.10~(-1)～3.10~(-3)g/ml人参蜂王浆后,细胞呈融合生长,认为人参蜂王浆对骨髓和脾细胞体外培养形成的小型CFU-F有成集落刺激作用,而对大型集落则相反。