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Giovanna Lacava Fulvio Laus Andrea Amaroli Andrea Marchegiani Roberta Censi Piera Di Martino Toru Yanagawa Maria Giovanna Sabbieti Dimitrios Agas 《Journal of cellular physiology》2019,234(9):16338-16347
With advancing age have been observed bone and bone marrow phenotypic alterations due to the impaired bone tissue homeostatic features, involving bone remodeling, and bone marrow niche ontogeny. The complex “inflamm-aging” pathological scenario that culminates with osteopenia and mesenchymal/stromal and hematopoietic stem cell commitment breakdown, is controlled by cellular and molecular intramural components comprising adapter proteins such as the sequestosome 1 (p62/SQSTM1). p62, a “multiway function” protein, has been reported as an effective anti-inflammatory, bone-building factor. In this view, we considered for the first time the involvement of p62 in aging bone and bone marrow of 1 year and 2 years p62−/− mice. Interestingly, p62 deficiency provoked accelerated osteopenia and impaired niche operational activities within the bone marrow. The above findings unearthed the importance of p62 in mesenchymal stem cell maintenance/differentiation schedule in old animals and provide, at least in part, a mechanistic scenario of p62 action. 相似文献
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Adipose cells have been recognized as an integral component of the bone marrow hematopoietic microenvironment in vivo and as an essential cell type required for in vitro maintenance of stem cells. Four stromal cell lines obtained from the adherent cell population of murine bone marrow cultures have been enriched and purified by multiple trypsinizations. We noted that these cell lines exhibited an accumulation of vacuoles of lipid, the extent of which varied be-tween cell lines in response to a change from medium containing 10% fetal calf serum to medium containing 20% horse serum. The lipid was lost when the cell lines were transferred back into the medium supplemented with fetal calf serum. In light of the reported lipogenic and antilipolytic effects of insulin on fibroblasts and adipocytes, we investigated the ability of insulin to induce adipocyte transformation of these bone marrow stromal cell populations. Three cell lines were exposed to bovine insulin at concentrations ranging from 10?9 to 10?6 M. All three cell lines responded to the insulin by accumulating lipid, but the extent of accumulation and the insulin concentration at which maximum lipid content was attained were population specific. One cell line (MC1) responded fully at physiological levels of insulin (10?9 M), whereas the other two showed lipid accumulation only at pharmacological concentrations. The initial growth of MC1 was inhibited in the presence of 10?9 M insulin which is compatible with the observed differentiation to adipocytes. The growth of MC3 was unaltered in the presence of physiological concentrations of insulin, whereas that of MC4 was accelerated. Grafts of organ cultures of the cell lines under the kidney capsule of syngeneic mice developed specific characteristics rep-resentative of the different cell lines. In particular, the majority of the grafts of MC1 consisted primarily of fat cells which were not observed in the grafts of MC3 and MC4. These data strongly suggest that these cell lines comprise cells with different potentialities and that the MC1 line represents a preadipocyte stromal cell of bone marrow. 相似文献
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When mesenchymal precursor cells from bone marrow are cultured in the presence of dexamethasone, the existence of distinct non-adherent and adherent populations can be demonstrated. The addition of PGE2, forskolin, or dibutyryl-cAMP can induce a transition from the former to the latter and this may be an important mechanism in the bone anabolic effects of PGE2. On the other hand, phorbol 12-myristate 13-acetate (PMA), an activator of protein kinase C, and sulprostone, an agonist for the PGE2 receptor EP1/EP3 subtypes, had no effect. The phosphodiesterase inhibitor, isobutylmethylxanthine (IBMX), had a synergistic effect in combination with PGE2, whereas neomycin, an inhibitor of inositol phosphate activity, had no effect, and LiCl, an inhibitor of inositol triphosphate metabolism, had an inhibitory effect on the PGE2-induced transition. Consistent with this, the addition of PGE2 to non-adherent bone marrow cells caused a 100% increase in cAMP synthesis. These results suggest that the induction of the transition from non-adherent to adherent osteoblast precursor is mediated by the EP2-PGE2 receptor subtype via an increase in intracellular cAMP synthesis. 相似文献
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人骨髓基质细胞体外分离及定向培养内皮细胞 总被引:2,自引:0,他引:2
用Ficoll(比重1.077 g/ml)从正常成人骨髓中分离骨髓基质细胞(BMSCs),DMEM-HG 培养基内含20?S、GM-CSF(100 u/ml)、VEGF(10 ng/ml)、FGF(5 ng/ml)、L-谷氨酰胺(2mmol/ L)、肝素(90 u/ml),以及抗生素液进行定向培养和扩增其中的内皮细胞(ECs),Ⅷ因子相关抗原的免疫组化法和透射电镜观察(TEM)鉴定其细胞的性质。结果5.0×105个BMSCs在体外经定向ECs 培养和扩增8代后,获得了6.0×109个ECs,扩增了约1.2×104倍。70%-80%的细胞对Ⅷ因子相关抗原免疫组化呈阳性反应;光镜下细胞呈典型的“鹅卵石”样;TEM下可观察到胞浆内有Weible- palade小体,证实为内皮细胞。实验表明,BMSCs在体外分离和定向培养的ECs,经扩增后可能是心血管组织工程所需种子细胞的又一个重要来源。 相似文献
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武妍周罗刘瑞琪黎明 《现代生物医学进展》2011,11(2):368-370
孕期应激对子代产生的影响是多方面的,这种影响是复杂的。研究表明,出生前的应激经历可导致出生后子代长期的免疫功能改变。这些改变追其根源与骨髓淋巴干细胞的改变有关。本文综述了大鼠孕期经历应激的子代骨髓淋巴干细胞所受的影响及免疫系统的相关改变,并根据现有的研究提出假说,为进一步研究孕期应激导致子代免疫系统改变的机理研究提供新的思路。 相似文献
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In Vitro Cellular &; Developmental Biology - Plant - 相似文献
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Mohammadhossein Hassanshahi Yu-Wen Su Chia-Ming Fan Samira Khabbazi Alireza Hassanshahi Cory J. Xian 《Journal of cellular biochemistry》2019,120(3):3220-3231
Chemotherapeutic agents are very well evident extrinsic stimuli for causing damage to endothelial cells. Methotrexate is an antimetabolite commonly used to treat solid tumours and paediatric cancers. However, studies on the effect(s) of methotrexate on bone marrow microvascular system are inadequate. In the current study, we observed a significant bone marrow microvascular dilation following methotrexate therapy in rats, accompanied by apoptosis induction in bone marrow sinusoidal endothelial cells, and followed by recovery of bone marrow sinusoids associated with increased proliferation of remaining bone marrow sinusoidal endothelial cells. Our in vitro studies revealed that methotrexate is cytotoxic for cultured sinusoidal endothelial cells and can also induce apoptosis which is associated with upregulation of expression ratio of Bax and Bcl-2 genes and Bax/Bcl-2 expression ratio. Furthermore, it was shown that methotrexate can negatively affect proliferation of cultured sinusoidal endothelial cells and also inhibit their abilities of migration and formation of microvessel like tubes. The data from this study indicates that methotrexate can cause significant bone marrow sinusoidal endothelium damage in vivo and induce apoptosis and inhibit proliferation, migration and tube-forming abilities of sinusoidal endothelial cells in vitro. 相似文献
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Antioxidants are known to influence metabolism and promote cell survival in a number of cell culture systems. However, their effects on the modulation of bone cell differentiationin vitroare not clearly defined. In the present studies we have investigated the effects of β-mercaptoethanol (βME) and ascorbate alone and in combination on human osteoprogenitors derived from bone marrow fibroblasts. In primary marrow cultures, βME stimulated colony formation (2-fold), alkaline phosphatase activity (3.5-fold) and, increased DNA synthesis (8-fold) after 21 days. Cell proliferation was increased significantly by βME during the first 4 days of a 10-day culture period, indicating stimulation of marrow osteoprogenitor proliferation. Ascorbate did not significantly augment the effects of βME in primary cultures or long-term cultures of passaged bone marrow fibroblasts. These findings indicate a potential beneficial role for βME addition for the optimal maintenance of colony formation, cell proliferation and differentiation of marrow osteoprogenitor cells in primary human bone marrow fibroblast cultures. 相似文献
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Tumor necrosis factor-alpha mediates RANK ligand stimulation of osteoclast differentiation by an autocrine mechanism 总被引:9,自引:0,他引:9
Osteoblasts or bone marrow stromal cells are required as supporting cells for the in vitro differentiation of osteoclasts from their progenitor cells. Soluble receptor activator of nuclear factor-kappaB ligand (RANKL) in the presence of macrophage colony-stimulating factor (M-CSF) is capable of replacing the supporting cells in promoting osteoclastogenesis. In the present study, using Balb/c-derived cultures, osteoclast formation in both systems-osteoblast/bone-marrow cell co-cultures and in RANKL-induced osteoclastogenesis-was inhibited by antibody to tumor necrosis factor-alpha (TNF-alpha), and was enhanced by the addition of this cytokine. TNF-alpha itself promoted osteoclastogenesis in the presence of M-CSF. However, even at high concentrations of TNF-alpha the efficiency of this activity was much lower than the osteoclastogenic activity of RANKL. RANKL increased the level of TNF-alpha mRNA and induced TNF-alpha release from osteoclast progenitors. Furthermore, antibody to p55 TNF-alpha receptors (TNF receptors-1) (but not to p75 TNF-alpha receptors (TNF receptors-2) inhibited effectively RANKL- (and TNF-alpha() induced osteoclastogenesis. Anti-TNF receptors-1 antibody failed to inhibit osteoclastogenesis in C57BL/6-derived cultures. Taken together, our data support the hypothesis that in Balb/c, but not in C57BL/6 (strains known to differ in inflammatory responses and cytokine modulation), TNF-alpha is an autocrine factor in osteoclasts, promoting their differentiation, and mediates, at least in part, RANKL's induction of osteoclastogenesis. 相似文献
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I. -D. Adler S. M. Attia 《Mutation Research - Genetic Toxicology and Environmental Mutagenesis》2003,542(1-2):139-142
Nicotne has been tested in the conventional mouse bone marrow assay. Single doses of 1 mg/kg bw or 2 mg/kg bw were given by oral intubations and bone marrow was sampled at 24 h (1 mg/kg) or at 6, 12 and 18 h after treatment (2 mg/kg). Nicotine treatment did not increase the micronucleus frequencies in polychromatic erythrocytes while the positive control compound mitomycin C yielded the expected result. These data contradict the only published in vivo study of nicotine in which 1.1 mg/kg bw was called positive for the induction of chromosomal aberrations in mouse bone marrow cells at all sampling intervals, even as early as 6 h after treatment. It is discussed that aberration scoring is a matter of subjectivity and depends on strict discrimination criteria between gaps and true DNA discontinuities, i.e. breaks. International collaboration has shown that micronucleus scoring is less subjective, hence more reliable. Therefore it is concluded that nocotine is not clastogenic at the doses and time intervals tested in the present experiments. 相似文献
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MBA-15, a marrow stromal-derived cell line, was shown to express an estrogen receptor. This finding was confirmed byin situhybridization and receptor binding assay. An exposure to estrogen (10?12–10?6m ) in a dose response manner resulted in a decrease of cell proliferation as measured by MTT assay. Cell function was measured by enzymatic activities of two osteoblastic markers, CD10/NEP and alkaline phosphatase. These enzymatic activities were elevated following the estrogen treatment. This model enabled direct evaluation of the estrogen effect on stromal osteoblast cells. 相似文献
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Bone Marrow Transplantation (BMT) is currently the most physiologic treatment for some types of leukodystrophies. In enzyme deficiency states, replacement of defective genes with cells carrying normal copies of these genes offers a natural form of gene therapy. This review will cover the various disease states which may be treated using bone marrow transplantation as well as the obstacles and advantages offered by this treatment modality. The potential for mixed hematopoietic chimerism, with reference to the advantages and disadvantages of treating various leukodystrophies, is reviewed. Finally, certain approaches which would reduce the morbidity and mortality associated with conventional BMT are discussed. If these obstacles can be overcome, BMT may offer the hope of cure to a number, but certainly not all, leukodystrophies. 相似文献
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低温冻存对骨髓基质细胞生物学特性的影响 总被引:4,自引:1,他引:3
目的:探讨低温冻存对骨髓细胞和贴壁基质细胞生物学特性的影响。方法:取新鲜骨髓和经Dexter法培养14d的骨髓贴壁基质细胞(称“基质细胞”),经-196℃液氮冻存(前者称“冻存骨髓”,后者称“冻存基质细胞”)2周,复温,再用Dexter法培养这些细胞,检测细胞增殖、细胞形态、细胞化学染色、细胞表面抗原及基质细胞支持另一骨髓造血细胞形成的鹅卵石造血区(CAFC),长期培养起始细胞(LTCIC)的变化,比较冻存对骨髓细胞和基质细胞生物学特性的影响。结果:生长特性:冻存骨髓比新鲜骨髓、冻存基质细胞比新鲜基质细胞培养后融合成片的时间延迟,细胞增殖数比也有减低。细胞成分:冻存骨髓比新鲜骨髓形成的成纤维细胞、内皮细胞比率下降,而巨噬细胞和脂肪细胞比率升高,冻存基质细胞上述现象更明显:冻存后含凋亡小体的细胞在骨髓细胞和基质细胞内均有增加。细胞表面抗原:冻存骨髓、冻存基质细胞CD14、HLA-DR抗原表达百分率比新鲜骨髓、新鲜基质细胞高,CD45、CD33反之。支持造血:冻存前后骨髓和基质细胞支持形成的CAFC和LTC-IC,生长良好,无显著差异。结论:骨髓细胞和经培养生成的贴壁基质细胞,经冻存和复温,生物学特性有一定变化,但仍可以保留良好的支持造血重建功能。 相似文献
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多发性骨髓瘤的骨髓细胞置于Dexter长期培养体系中,浆细胞可以长期生长。非贴壁层细胞形成CFU-GM的能力达6周以上,但随着培养时间的延长,CFU-GM的形成活性明显下降。浆细胞产生兔疫球蛋白单克隆特异性在Dexter长期培养体系中仍维持不变,但Ki-67阳性的浆细胞随着培养时间的延长而减少,以上结果说明浆细胞虽然可以在Dexter培养体系中生长并保持单克隆特异性,但此种培养体系不是浆细胞最佳的生长条件。本研究还表明Dexter长期培养不宜作为骨髓净化的手段用于多发性骨髓瘤的自体骨髓移植。 相似文献
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《Cell reports》2020,30(4):949-958.e6
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Zinc and fluorides are capable of modifying the process of bone formation and mineralization. Statistically significant differences
have been revealed in the content of zinc and fluorides between structures of the femur in chicken. The content of zinc in
compact bone remained constant during the first 50 d of life. Lower and less stable contents were found in spongy bone and
bone marrow. The content of fluorides in compact bone was higher than in spongy bone. The lowest concentrations of zinc and
fluorides were found in articular cartilage and were further reduced at the end of observation. Correlations revealed between
the content of zinc and fluorides point to structural and functional relationships between these elements in various parts
of the bone. 相似文献