草鱼肾脏和脾脏血细胞发育过程的观察

鱼类肾脏、脾脏是机体造血的主要器官。印迹涂片显示:草鱼肾脏和脾脏内血细胞的发育过程大致经历了三个阶段,即原始阶段、幼稚阶段、成熟阶段。本文着重描述了各阶段细胞的形态特征井对草鱼血细胞的发育及命名等问题作了初步的探讨。       

造血干细胞发生学研究 Ⅲ.人胚脾和胸腺造血

前已报道人胚肝、骨髓造血的研究资料,对于该两组织造血发育的规律,实质细胞和间质细胞的相互联系,造血干细胞的可能来源等问题提出了一些设想和实验依据。而脾脏和胸腺在胚胎时期的一定阶段亦是造血功能的执行者,在这方面文献中涉及甚少,尤其是有关电子显微镜的资料更为缺乏。本文对此两种胚胎组织进行光镜和透射电镜的观察,以     

中华鳖造血和免疫器官的个体发育

采用常规孵化的中华鳖胚胎为材料,对不同发育时期造血和免疫器官进行了组织学研究,描述了卵黄囊、胸腺、肝、脾、肾以及骨髓的形态结构变化。发现胚胎期首先出现的造血器官是卵黄囊。此后,卵黄囊的造血干细胞出现在胚体的血循环中,造血功能相继在胚胎胸腺、肝、脾、骨髓(可能还包括肾)中产生。胸腺是中华鳖免疫系统发育的第一个淋巴器官,来自卵黄囊的干细胞在此先分化成小淋巴细胞,然后再迁移至脾脏。脾脏发育中首先出现各发育阶段的红细胞、嗜酸性的细胞和少量粒细胞,淋巴细胞出现较晚,未发现淋巴小结。在胚胎期肝脏发育过程中可见不同发育时期的红细胞和嗜酸性的细胞。在肾的发育过程中,尚可观察到嗜酸性的细胞和类似头肾组织的细胞团。直至出壳前,骨髓内方可见各发育阶段的各系细胞[动物学报49(2)：238—247,2003]。     

长吻鮠血细胞发生的研究

用Wright-Giemsa和PAS染色对长吻鮠头肾、肾脏、脾脏、肝脏等器官组织的涂片、印片染色观察发现,头肾、肾脏和脾脏是其主要造血器官。红细胞、粒细胞和淋巴细胞主要在肾脏和头肾中发生,其次是脾脏。单核细胞则主要在肾脏和脾脏中发生,头肾中也有少量单核细胞产生。肝脏中无原始型血细胞,可能不是其造血器官。红细胞的发育经历四个阶段,其胞体体积经历了由大到小,由小到大再变小的"两大两小"发育过程;粒细胞的发育经历五个阶段,其胞体体积均由大变小,双叶或多叶核的粒细胞可能是衰老的粒细胞亦即核的分叶是粒细胞衰老的标志;淋巴细胞和单核细胞的发育各经历了三个阶段,两者发育成熟过程中胞体体积均由大变小。巨噬细胞由单核细胞发育而来。原血细胞和部分早期幼稚血细胞可以进行有丝分裂,部分成熟红细胞和血栓细胞可以进行直接分裂。红细胞在整个发育过程中,PAS反应均呈阴性,各类白细胞的发育过程中,PAS反应由阴性到阳性并逐渐增强,这显示随着白细胞的逐渐发育成熟,细胞内糖原物质含量逐渐增多。     

长吻(鱼危)血细胞发生的研究

用Wright-Giemsa和PAS染色对长吻(鱼危)头肾、肾脏、脾脏、肝脏等器官组织的涂片、印片染色观察发现,头肾、肾脏和脾脏是其主要造血器官.红细胞、粒细胞和淋巴细胞主要在肾脏和头肾中发生,其次是脾脏.单核细胞则主要在肾脏和脾脏中发生,头肾中也有少量单核细胞产生.肝脏中无原始型血细胞,可能不是其造血器官.红细胞的发育经历四个阶段,其胞体体积经历了由大到小,由小到大再变小的"两大两小"发育过程;粒细胞的发育经历五个阶段,其胞体体积均由大变小,双叶或多叶核的粒细胞可能是衰老的粒细胞亦即核的分叶是粒细胞衰老的标志;淋巴细胞和单核细胞的发育各经历了三个阶段,两者发育成熟过程中胞体体积均由大变小.巨噬细胞由单核细胞发育而来.原血细胞和部分早期幼稚血细胞可以进行有丝分裂,部分成熟红细胞和血栓细胞可以进行直接分裂.红细胞在整个发育过程中,PAS反应均呈阴性,各类白细胞的发育过程中,PAS反应由阴性到阳性并逐渐增强,这显示随着白细胞的逐渐发育成熟,细胞内糖原物质含量逐渐增多.     

WIP1对小鼠B细胞及T细胞发育的影响

目的研究WIP1基因对小鼠骨髓B细胞发育及胸腺T细胞发育的影响。方法流式细胞术测定小鼠骨髓B细胞及胸腺T细胞发育中各阶段的细胞比例。结果虽然WIP1缺失小鼠骨髓B细胞发育各阶段比例正常,但骨髓总体B细胞比例下降;WIP1基因敲除小鼠胸腺发育障碍,CD8/CD4双阴性细胞比例增高,CD8/CD4双阳性细胞比例降低。结论 WIP1基因在小鼠骨髓B细胞及胸腺T细胞的发育过程中起重要作用。     

Cramp过表达对小鼠骨髓造血干细胞功能的影响

目的研究Cramp蛋白过表达对小鼠骨髓造血干细胞自我更新和分化能力的影响。方法应用流式细胞仪分析Cramp过表达转基因小鼠及同龄野生型小鼠的骨髓、脾脏、胸腺等组织器官中各种细胞的比例;分选骨髓造血干细胞,体外培养,观察其克隆形成能力。结果与野生型小鼠相比,Cramp过表达转基因小鼠的骨髓、脾脏、胸腺等组织器官中各种细胞的比例、骨髓造血干细胞的克隆形成能力等均无明显变化。结论本研究中,Cramp过表达转基因小鼠骨髓造血干细胞的分化能力、克隆形成能力无明显变化。     

小鼠体内脾脏,骨髓及精原细胞姐妹染色单体交换的比较研究

本文对几种化学诱变剂诱发小鼠体内脾脏、骨髓和精原细胞的SCE进行了比较研究,同时分析了几类常见化合物在小鼠脾脏细胞中诱发SCE的活力。结果显示诱变剂在脾脏细胞中诱发SCE比骨髓和精原细胞敏感。几类化合物都能显著地诱发小鼠脾脏SCE的增加,与对照相比差异显著(P<0.05)或极显著(P<0.01),说明利用小鼠脾脏细胞检测环境诱变物是相当灵敏的。     

乌龟胚胎发育的研究

根据乌龟胚胎发育过程中形态特征和内部结构的变化,将其胚胎发育从卵产出到幼龟孵出划分为28期。分别描述了各期的主要特征。并就乌龟发育分期的标准,原肠胚的形成,羊膜尾褶和后羊膜管,早期胚胎的自身运动等问题进行了讨论。     

胸腺对法氏囊滤泡及脾脏B淋巴细胞区发育的影响

Siskind 等(1979)发现胸腺或胸腺细胞对B细胞接触抗原前的发育有影响;未成熟B细胞对胸腺依赖性抗原反应而产生异质性抗体的能力受胸腺的控制。Bhogal 等(1984)进一步指出,鸡法氏囊 B细胞的发育存在一个胸腺依赖的阶段,胸腺对法氏囊的发育可能存在体液性影响。本文通过手术去除胸腺、注射胸腺提取液等实验,观察法氏囊和脾脏淋巴细胞的显微和亚显微结构的变化,探讨胸腺对法氏囊滤泡及脾脏 B细胞区发育的影响。 材料和方法:实验用初生雏鸡是从山东省农科院购买的莱杭雏鸡,常规饲养。实验每组 10只雏鸡,分以下几组进行:1.手术切除胸腺组,取刚孵出的雏鸡,手术去除胸腺;2.手术切除胸腺并注射同龄鸡     

乌龟脑垂体显微及其腺垂体超微结构的研究

乌龟脑垂体由柄形神经垂体和椭圆形腺垂体两部分组成,神经垂体位于腺垂体后部上方呈背腹型排列。神经垂体中神经叶不发达,腺垂体分为远侧部和中间部,特殊空泡结构成为垂体门脉系统的特征。远侧部细胞分为嗜酸性细胞、嗜碱性细胞和嫌色细胞3种。通过透射电镜观察,腺垂体远侧部主要有5种分泌激素细胞:即生长激素(GH)分泌细胞、催乳激素(PRL)分泌细胞、促甲状腺激素(TSH)分泌细胞、促肾上腺皮质激素(ACTH)分泌细胞、促性腺激素(GTH)分泌细胞和非分泌类型滤泡-星形细胞(FS)。生长激素分泌细胞核大、分泌颗粒少的特征成为乌龟与其他动物最大的区别,可能与乌龟具有生长慢、寿命长的生物学特性有关。       

乌脑龟垂体显微及其腺垂体超微结构的研究

乌龟脑垂体由柄形神经垂体和椭圆形腺垂体两部分组成,神经垂体位于腺垂体后部上方呈背腹型排列。神经垂体中神经叶不发达,腺垂体分为远侧部和中间部,特殊空泡结构成为垂体门脉系统的特征。远侧部细胞分为嗜酸性细胞、嗜碱性细胞和嫌色细胞3种。通过透射电镜观察,腺垂体远侧部主要有5种分泌激素细胞：即生长激素（GH）分泌细胞、催乳激素（PRL）分泌细胞、促甲状腺激素（TSH）分泌细胞、促肾上腺皮质激素（ACTH）分泌细胞、促性腺激素（GTH）分泌细胞和非分泌类型滤泡．星形细胞（Fs）。生长激素分泌细胞核大、分泌颗粒少的特征成为乌龟与其他动物最大的区别,可能与乌龟具有生长慢、寿命长的生物学特性有关。     

Antibody formation in mouse bone marrow. VIII. Dependence on potentially circulating memory cells: a study with parabiotic mice.

Mouse bone marrow barely contains antibody-producing plaque-forming cells (PFC) during the primary response to sheep red blood cells (SRBC). However, during the secondary response, the number of IgM, IgG, and IgA PFC in the bone marrow can rise to a level which surpasses the number of PFC in all the other lymphoid organs together. In the present paper we investigated whether the capacity of immune mice to react upon a booster injection of SRBC with a bone marrow PFC response can be transferred from immune to nonimmune mice. Therefore, mice primed with SRBC 6 months previously and nonprimed syngeneic mice were joined for parabiosis and were separated from each other at various intervals after joining. These separated mice were subsequently immunized with SRBC. It was found that, after 3 weeks of parabiosis, the nonprimed members reacted upon an injection of SRBC with a bone marrow IgM, IgG, and IgA PFC response as high as did the previously primed members. Furthermore it could be demonstrated by means of cell transfer experiments that, after a period of parabiosis of 3 weeks, the bone marrow and spleen of the normal mice contained about as many memory cells as the bone marrow and spleen of the immune mice. These results suggest that antibody formation in mouse bone marrow is dependent on a population of potentially circulating memory cells.     

Mechanisms of lymphocytic choriomeningitis virus-induced hemopoietic dysfunction.

Results of this study showed that lymphocytic choriomeningitis virus infection causes a marked activation of natural killer (NK) cells not only in the spleen but also in the bone marrow. This activity reached its peak at about day 3 of infection and declined after days 6 to 7. Enhanced NK cell activity was found to correlate with decreased receptivity for syngeneic stem cells in bone marrow and spleen, with the notable exception that decreased receptivity persisted longer in bone marrow. Treatment of infected recipients with anti-asialo GM1 (ganglio-N-tetraosylceramide) significantly increased the receptivity for syngeneic hemopoietic cells. These findings are consistent with the hypothesis that NK cell activation causes rejection of syngeneic stem cells, thus resulting in hemopoietic depression. To understand the mechanisms behind the prolonged decrease in bone marrow receptivity (and bone marrow function in the intact mouse) mentioned above, we followed the changes in the number of pluripotential stem cells (CFU-S) circulating in the peripheral blood and in endogenous spleen colonies in irradiated mice, the limbs of which were partially shielded. It was found that following a marked early decline, both parameters increased to normal or supranormal levels at about day 9 after infection. Because the bone marrow pool of CFU-S is only about 20% of normal at this time after infection, a marked tendency for CFU-S at this stage in the infection to migrate from the bone marrow to the spleen is suggested. It seems, therefore, that as NK cell activity declines, the spleen regains the ability to support growth of hemopoietic cells and the bone marrow resumes an elevated export of stem cells to the spleen. This diversion of hemopoiesis could explain both the long-standing deficiencies of the bone marrow compartment and the prolonged decrease in the receptivity of this organ.     

Exonuclease-1缺失延缓端粒酶缺失小鼠造血微环境的衰老

目的研究Exo-1对端粒酶缺失小鼠造血微环境衰老的影响。方法以端粒酶基因敲除小鼠（Terc-/-）和Exo-1基因敲除小鼠（Exo-1-/-）杂交,并进一步互交产生第三代端粒酶基因敲除小鼠（G3Terc-/-）以及第三代Terc和Exo-1双基因敲除小鼠（G3Terc-/-Exo-1-/-）。以CD45.1野生型小鼠的骨髓细胞为供体,以2月龄G3Terc-/-或G3Terc-/-Exo-1-/-小鼠为受体,进行骨髓移植。在受体小鼠9月龄时,取骨髓、脾脏、胸腺、外周血等组织器官的细胞进行流式分析,研究G3Terc-/-和G3Terc-/-Exo-1-/-受体小鼠中的野生型供体来源的造血干细胞的发育分化。结果同G3Terc-/-小鼠相比,G3Terc-/-Exo-1-/-双基因敲除受体小鼠骨髓中野生型供体来源的B220＋细胞比例升高,前体B细胞的比例也明显升高;脾脏B220＋细胞的比例明显升高;胸腺发育正常;外周血中B220＋细胞比例升高。结论 Exo-1缺失延缓了端粒酶基因敲除小鼠造血系统微环境的衰老,从而逆转了端粒功能障碍引起的骨髓造血干细胞发育分化异常。     

Cytokine-pretreatment of CD34(+) cord blood stem cells in vitro reduces long-term cell engraftment in NOD/SCID mice

Stem cell homing, engraftment and organ regeneration are controlled by cytokines, chemokines and cell-cell interactions. In this paper, cytokine effects on homing- and engraftment-related characteristics of CD34(+) cord blood cells were examined. Untreated CD34(+) cells were mainly in the G(0)/G(1) cell cycle phase, expressed adhesion receptors on a low level, were positive for vimentin, and negative for the epithelial marker cytokeratin 8/18. Treatment with stem cell factor (SCF) stimulated cell proliferation, increased the number of cells in S and G(2)/M cell cycle phase as well as the expression of adhesion receptors. The expression of cytokeratin 8/18 was increased and that of vimentin remained unchanged. Hepatocyte growth factor (HGF) did not stimulate cell proliferation and expression of adhesion receptors, but increased expression of cytokeratin 8/18. In NOD/SCID mice, kinetics of stem cell distribution revealed a fast elimination of human cells from blood. An increase in the number of engrafted cells was observed in different mouse organs in a time-dependent manner, preferentially in bone marrow, spleen and liver. Pretreatment with SCF resulted in reduction of long-term engraftment in bone marrow. HGF pretreatment of cord blood cells showed no significant effects on long-term engraftment capacity in mouse organs compared to untreated cells. Our data provide in vivo evidence that pretreatment of CD34(+) cells with SCF reduces long-term cell engraftment in NOD/SCID mice.     

Unbiased selection of bone marrow derived cells as carriers for cancer gene therapy

BACKGROUND: There is currently great interest in development of cell-based carriers for delivery of viral vectors to metastatic tumors. To date, several cell carriers have been tested based largely upon their predicted tumor-localizing properties. However, cell types may exist which can be mobilized from the circulation by a tumor which have not yet been identified. Here we use an unbiased screen of bone marrow (BM) cells to identify cells which localize to tumors and which might serve as effective candidate cell carriers without any prior prediction or selection. METHODS: Unsorted BM cells from green fluorescent protein (GFP)-transgenic donor mice were adoptively transferred into C57Bl/6 mice bearing pre-established subcutaneous B16 melanoma tumors. Forty-eight hours and eight days later, tumors, organs and blood were analyzed for GFP-expressing cells by flow cytometry. The phenotype of GFP cells in organs was determined by co-staining with specific cell surface markers. RESULTS: CD45(+) hematopoietic cells were readily detected in tumor, spleen, bone marrow, blood and lung at both time points. Within these CD45(+) cell populations, preferential accumulation in the tumor was observed of cells expressing Sca-1, c-kit, NK1.1, Thy1.2, CD14, Mac-3 and/or CD11c. Lymphodepletion increased homing to spleen and bone marrow, but not to tumors. CONCLUSIONS: We have used an in vivo screen to identify populations of BM-derived donor cells which accumulate within tumors. These studies will direct rational selection of specific cell types which can be tested in standardized assays of cell carrier efficiency for the treatment of metastatic tumors.     

A model of Ph' positive chronic myeloid leukemia-blast crisis cell line growth in immunodeficient SCID mice.

A human Philadelphia-chromosome positive chronic myeloid leukemia-blast crisis (CML-BC) cell line BV173 proliferated in the hematopoietic tissues, infiltrated various organs and caused the death of immunodeficient SCID mice. Leukemia spreading was assessed with diminished number of bone marrow cells and caused splenomegaly. The leukemic colonies grew from single cell suspension of bone marrow, spleen and peripheral blood. Bcr-abl m-RNA was detectable in bone marrow, spleen, peripheral blood, liver, lungs and brain. Dying mice demonstrated severely hypoplastic bone marrow, splenomegaly and massive metastases in the liver and kidneys. The survival time of animals was dependent on the number of inoculated leukemia cells.