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1.
Identification and characterization of osteoclast-like cells and their progenitors in cultures of feline marrow mononuclear cells 总被引:11,自引:1,他引:10
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《The Journal of cell biology》1984,99(2):471-480
The predominant cell responsible for bone resorption, the multinucleated osteoclast, has been difficult to study because of inaccessibility. When feline marrow-derived mononuclear cells are established in long-term culture, multinucleated cells form within 48 h, reaching maximum numbers at 16 d. We have observed that these cultured cells have many of the features of osteoclasts. Morphologically, they are multinucleated, contain large numbers of branched mitochondria, have a peripheral cytoplasm lacking organelles (a clear zone), and have extensive cell-surface processes. In addition to these ultrastructural features, the cells contain a tartrate- resistant acid phosphatase, the activity of which is increased by parathyroid hormone (PTH) and inhibited by calcitonin. PTH, prostaglandin E2, and 1,25(OH)2 vitamin D3 increased multinucleated cell formation, while calcitonin inhibited the stimulatory effects of PTH. Time-lapse cinemicrographic and autoradiographic studies indicated that the multinucleated cells formed by fusion of the mononuclear progenitors. The multinucleated cells were phagocytic and stained with nonspecific esterase, consistent with their being derived from immature monocytes. Further, cell populations enriched for multinucleated cells release 45Ca from devitalized bone. Density-gradient centrifugation on Percoll was used to enrich and characterize the mononuclear progenitors of these multinucleated cells. The progenitor cells were found predominantly in Percoll density layers of 1.065 to 1.08 g/ml and were enriched up to 30-fold as compared to unfractionated cells. The bone marrow mononuclear cells that formed the multinucleated cells were initially nonadherent to plastic, stained heavily with nonspecific esterase, and appeared to be immature monocytes histologically. These data indicate that the multinucleated osteoclast-like cells in our cultures are derived from nonadherent monocytic progenitor cells that are responsive to osteotropic hormones. The ability to grow and characterize these cells in vitro should facilitate studies to elucidate the role these cells play in normal and pathologic states of bone resorption. 相似文献
2.
H P Baum W Thoenes 《Virchows Archiv. B, Cell pathology including molecular pathology》1985,50(2):181-192
Morphometric analysis disclosed distinct differences between blood monocytes, tissue monocytes (i.e. immature macrophages), epithelioid cells and multinucleated giant cells as well as phagocytic macrophages (i.e. mature macrophages) in the granuloma model of autoimmune (anti-TBM) tubulo-interstitial nephritis. The numerical density of lysosomes decreased slightly in tissue monocytes compared with blood monocytes but showed a pronounced increase during the formation of epithelioid cells. The lysosomal compartments of epithelioid cells and multinucleated giant cells resembled each other very closely, but the giant cells obviously produced additional lysosomes of small diameter (80-120 nm). Phagocytic macrophages displayed a total numerical density of lysosomes similar to that of tissue monocytes but the mean diameter of the lysosomes was markedly greater. Thus the volume density of lysosomes was highest in phagocytic macrophages. The blood monocytes exhibited the smallest lysosomal compartment. In tissue monocytes, epithelioid cells, and multinucleated giant cells the volume densities of the lysosomes were greater than in blood monocytes and remained relatively constant because the increase in numerical density was counterbalanced by a decrease in mean granule diameter. We found only minor differences in mitochondrial volume densities among the five cell populations. The shape of the mitochondria, however, changed steadily from short rotational ellipsoids in the blood monocytes to rather elongated and slender bodies in the multinucleated giant cells. The results suggest that epithelioid cells and multinucleated giant cells are active cells which may contribute by their specific performances, to the immunologic microenvironment of the granuloma. 相似文献
3.
Cell size and incidence of multinucleated, polyploid cells in cultured aortic smooth muscle cells from different age groups of spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto rats (WKY) were compared. Smooth muscle cells from SHR were generally larger than those from WKY, and the percentage of multinucleated smooth muscle cells was always higher in SHR than WKY in the three age groups of rats studied (3-4, 10-12, and 28-30 weeks). In smooth muscle cells from the 3- to 4-week group, there was a positive correlation between cell diameter and the percentage of multinucleated smooth muscle cells. Microdensitometric measurements also showed that the incidence of polyploid smooth muscle cells was always higher in SHR than WKY in the three age groups. There was a positive correlation between DNA density and nuclear area measurements in all the age groups of SHR and WKY. We conclude that cultured aortic smooth muscle cells from different age groups of SHR and WKY contained heterogeneous populations of cells and that, under our culture conditions, the polyploidy of the smooth muscle cells found in vivo was maintained in the SHR and WKY. 相似文献
4.
A Malassiné C Besse A Roche E Alsat R Rebourcet F Mondon L Cedard 《Histochemistry》1987,87(5):457-464
Low density lipoproteins (LDL) were conjugated to colloidal gold to visualize the route for internalization of LDL in the cultured cells of human term placenta. Cells were obtained from placental villi (caesarian section) by a standard trypsin-DNase dispersion method followed in some cases by a Percoll gradient centrifugation step. Employing electron microscopy it was observed that after 3 days of culture, cells obtained by trypsin-DNase dispersion were a mixture of macrophages, mononucleated cells and large multinucleated cells. When the cells were incubated for 3 days after the Percoll purification, essentially multinucleated cells identical to the syncytiotrophoblast were present. The number of LDL receptor was increased by preincubation in medium with lipoprotein depleted serum. In binding experiments cells incubated at 4 degrees C for 2 h with medium containing gold LDL conjugates showed gold LDL attached to the plasma membrane without characteristic localization. After incubation with gold LDL at 37 degrees C for various times, the three cellular types showed ligand internalization. Gold LDL endocytosis involved first coated pits but also uncoated plasmalemmal invaginations. Then gold LDL was further observed in coated and non coated vesicles, smooth walled endosomes, multivesicular bodies and tubular vesicles. Lastly free gold particles were observed in lysosome like dense bodies. These results prove the internalization of gold LDL conjugates by human cultured placental cells, particularly by syncytiotrophoblast like multinucleated cells. This accumulation of LDL (the major cholesterol carrying protein in humans) is recognised to be responsible for the exogenous cholesterol supply indispensable to the progesterone biosynthesis and cellular growth of the placenta. 相似文献
5.
A. Malassiné C. Besse A. Roche E. Alsat R. Rebourcet F. Mondon L. Cedard 《Histochemistry and cell biology》1987,87(5):457-464
Summary Low density lipoproteins (LDL) were conjugated to colloidal gold to visualize the route for internalization of LDL in the cultured cells of human term placenta. Cells were obtained from placental villi (caesarian section) by a standard trypsin-DNase dispersion method followed in some cases by a Percoll gradient centrifugation step. Employing electron microscopy it was observed that after 3 days of culture, cells obtained by trypsin-DNAse dispersion were a mixture of macrophages, mononucleated cells and large multinucleated cells. When the cells were incubated for 3 days after the Percoll purification, essentially multinucleated cells identical to the syncytiotrophoblast were present. The number of LDL receptor was increased by preincubation in medium with lipoprotein depleted serum. In binding experiments cells incubated at 4° C for 2 h with medium containing gold LDL conjugates showed gold LDL attached to the plasma membrane without characteristic localization. After incubation with gold LDL at 37° C for various times, the three cellular types showed ligand internalization. Gold LDL endocytosis involved first coated pits but also uncoated plasmalemmal invaginations. Then gold LDL was further observed in coated and non coated vesicles, smooth walled endosomes, multivesicular bodies and tubular vesicles. Lastly free gold particles were observed in lysosome like dense bodies. These results prove the internalization of gold LDL conjugates by human cultured placental cells, particularly by syncytiotrophoblast like multinucleated cells. This accumulation of LDL (the major cholesterol carrying protein in humans) is recognised to be responsable for the exogenous cholesterol supply indispensable to the progesterone biosynthesis and cellular growth of the placenta. 相似文献
6.
The cytomorphologic features were analyzed in thyroid samples obtained by fine needle aspiration (FNA) from 54 cases proven to be Grave's disease (toxic goiter) by a multiparameter study. A fire-flare appearance of the follicular cells and a good cellularity of the smears were the features most frequently observed in these cases. Hürthle cell changes were present in 47% of the cases, and small numbers of lymphocytes were seen in smears from 41% of the cases. Epithelioid cell granulomata and multinucleated giant cells were observed in less than one-fourth of the cases. About half of the cases showed some degree of pleomorphism of the follicular cells. Twenty cases of toxic multinodular goiter were similarly analyzed; the features that distinguished this entity from Grave's disease were the absence of epithelioid cells and multinucleated giant cells in the FNA smears. 相似文献
7.
We have previously shown that Elasmobranchs-characterized by a partially calcified cartilaginous endoskeleton-presented a bony vertebral arch containing osteoblasts, osteocytes and resorbing cells. The aim of this study is to test the ability of Elasmobranchs to resorb bone tissue. The subcutaneous implantation in dogfish (Scyliorhinus canicula) of devitalized mineral-containing bone particles, obtained from a bony fish (the eel, Anguilla anguilla) resulted, after 21 2 months, in the formation of mononucleated as well as multinucleated cells around and between the bone fragments. By light microscopy, the multinucleated giant cells presented the general aspect of osteoclastic cells whereas, by transmission electron microscopy they never showed ruffled borders which are considered as the typical features of osteoclasts. Except for this character, the mononucleated and multinucleated cells exhibited the typical ultrastructural aspects leading us to say that these cells are involved in the resorption of the bone fragments. This study shows that Elasmobranchs are able to resorb implanted bone. 相似文献
8.
Large scale gene expression analysis of osteoclastogenesis in vitro and elucidation of NFAT2 as a key regulator 总被引:19,自引:0,他引:19
Ishida N Hayashi K Hoshijima M Ogawa T Koga S Miyatake Y Kumegawa M Kimura T Takeya T 《The Journal of biological chemistry》2002,277(43):41147-41156
To understand the molecular events coupling between cell proliferation and differentiation by elucidating genes essential for the process, we conducted a large scale gene expression analysis of an in vitro osteoclastogenesis system consisting of recombinant RANKL and mouse RAW264 cells. The entire process leading to the formation of tartrate resistant acid phosphatase-positive multinucleated cells takes 3 days and plates become fully covered with multinucleated cells at 4 days. Microarray probing at eight time points revealed 635 genes that showed greater than 2-fold differential expression for at least one time point and they could be classified into six groups by the "k-means" clustering analysis. Among a group of 106 early inducible genes (within 2-5 h after RANKL stimulation), four genes including NFAT2 were identified as genes whose enhanced expressions were fairly correlated with an efficient induction of matured osteoclasts. Moreover, cyclosporin A significantly suppressed the multinucleated cell formation accompanying the reduction of the nuclear localization of NFAT2. When the expression of NFAT2 was suppressed by introducing antisense NFAT2, multinucleated cell formation was severely hampered. Functional analysis thus combined with gene analysis by microarray technology elucidated a key role of NFAT2 in osteoclastogenesis in vitro. 相似文献
9.
Jindrich Cinatl Jr. Jaroslav Cinatl Pablo Hernáiz Driever Holger Rabenau Milos Novak Milan Stefanik Bernhard Kornhuber Hans-Wilhelm Doerr 《Cell biology international》1994,18(4):271-278
The effects of aphidicolin, a specific inhibitor of DNA polymerase α, on cell growth, DNA synthesis and myogenic differentiation in the human alveolar rhabdomyosarcoma cell line KFR were studied. The treatment with aphidicolin at 5 × 10−6 M concentration, which completely inhibited DNA synthesis and cell growth, induced morphological differentiation of small mononuclear cells to elongated, multinucleated (myotube-like) structures. The morphological differentiation was accompanied by the expression of skeletal muscle myosin; about 30% myosin-positive cells were observed after 14 days of treatment, compared to 2.3% in untreated cultures. The results showed that aphidicolin induces differentiation of human rhabdomyosarcoma cells and that multinucleated myotube-like elements may develop simply by cell fusion without cell division and DNA synthesis. 相似文献
10.
Ng WK 《Acta cytologica》2002,46(3):535-539
OBJECTIVE: To describe the fine needle aspiration cytology findings of fibroadenoma with multinucleated stromal giant cells, with histologic correlation. STUDY DESIGN: The author reviewed the cytologic findings of two cases of fibroadenoma with multinucleated stromal giant cells from the file of Pamela Youde Nethersole Eastern Hospital in a six-year period from 1995 to the end of 2000. The diagnosis was confirmed by histologic examination of the lumpectomy specimens. RESULTS: The two cases had similar cytologic findings. The direct smears contained cohesive clusters of bland-looking ductal cells arranged in a "staghorn" pattern. Numerous naked nuclei were also seen in the background. Also, there were occasional multinucleated giant cells in isolation. These giant cells contained 5-10 randomly arranged, round to oval nuclei, fine chromatin and sometimes distinct nucleoli. The cytoplasm was abundant and pale staining, and the cell border was ill defined. Associated epithelioid histiocytes and foamy macrophages were not seen. Histologic examination of the lumpectomy specimens showed architectural features of fibroadenoma with pericanalicular and intracanalicular patterns. In addition, scattered multinucleated giant cells with focal degenerative change were noted in the tumor stroma. Their stromal nature was confirmed by immunohistochemical study. CONCLUSION: Multinucleated stromal giant cells are rarely identified in fine needle aspiration biopsies of fibroadenoma. Recognition of this peculiar finding may help to avoid misdiagnosis of other, more sinister conditions, such as phyllodes tumor and metaplastic carcinoma. 相似文献
11.
12.
About ScienceDirect 《Experimental cell research》1982,137(2):489-494
Satellite cells were isolated from leg skeletal muscles of adult mice and grown in culture. During the first few days in culture, satellite cells actively proliferated and starting on day 4 began to fuse into multinucleated myotubes. At various time points during the culture period, the biosynthesis of total cellular proteins and glycoproteins was analysed by pulse-labelling with radioactive leucine or sugars followed by electrophoretic analysis on two-dimensional gels. Our findings are: (1) Replicating mononucleated satellite cells on day 1 of culture did not synthesize detectable amounts of α and β tropomyosins, α-actin, and myosin light chains 1 and 2; (2) the synthesis of these polypeptides was readily detectable in multinucleated myotubes that formed by day 5–6 of culture; (3) other qualitative and quantitative differences in as yet unidentified proteins were observed in replicating cells as compared with multinucleated myotubes as well as to muscle fibroblasts; and (4) at least two distinct glucosamine-containing acidic glycoproteins of about 70 000 D and pI 5 were synthesized by myotubes, but not by replicating satellite cells.These data demonstrate that the biosynthetic programs for proteins and glycoproteins of cultured replicating satellite cells can be distinguished from those of multinucleated myotubes and from those of muscle fibroblasts. These data are interpreted to indicate that during muscle histogenesis in vivo, satellite cells become arrested prior to the expression of terminally differentiated traits. 相似文献
13.
Satellite cells were isolated from leg skeletal muscles of adult mice and grown in culture. During the first few days in culture, satellite cells actively proliferated and starting on day 4 began to fuse into multinucleated myotubes. At various time points during the culture period, the biosynthesis of total cellular proteins and glycoproteins was analysed by pulse-labelling with radioactive leucine or sugars followed by electrophoretic analysis on two-dimensional gels. Our findings are: (1) Replicating mononucleated satellite cells on day 1 of culture did not synthesize detectable amounts of α and β tropomyosins, α-actin, and myosin light chains 1 and 2; (2) the synthesis of these polypeptides was readily detectable in multinucleated myotubes that formed by day 5–6 of culture; (3) other qualitative and quantitative differences in as yet unidentified proteins were observed in replicating cells as compared with multinucleated myotubes as well as to muscle fibroblasts; and (4) at least two distinct glucosamine-containing acidic glycoproteins of about 70 000 D and pI 5 were synthesized by myotubes, but not by replicating satellite cells.These data demonstrate that the biosynthetic programs for proteins and glycoproteins of cultured replicating satellite cells can be distinguished from those of multinucleated myotubes and from those of muscle fibroblasts. These data are interpreted to indicate that during muscle histogenesis in vivo, satellite cells become arrested prior to the expression of terminally differentiated traits. 相似文献
14.
Local expression and exocytosis of viral glycoproteins in multinucleated muscle cells 总被引:3,自引:2,他引:1
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《The Journal of cell biology》1992,117(5):987-995
We have analyzed the distribution of enveloped viral infections in multinucleated L6 muscle cells. A temperature-sensitive vesicular stomatitis virus (mutant VSV ts045) was utilized at the nonpermissive temperature (39 degrees C). As expected, the glycoprotein (G protein) of this mutant was restricted to the ER when the multinucleated cells were maintained at 39 degrees C. We demonstrate that this G protein remained localized when the infection was performed at low dose. By 4 h after infection the G protein patches spanned an average of 220 microns. The localization was independent of nuclear positions, showing that the ER was a peripheric structure. Thus, the infection did not recognize nuclear domains characteristic of nuclearly encoded proteins. After release of the 39 degrees C block, transport through a perinuclear compartment into a restricted surface domain lying above the internal G protein patch occurred. Accordingly, the transport pathway was locally restricted. After a 16-h infection the G protein spanned 420 microns, while the matrix protein occupied 700-800 microns of the myotube length. Double infection of multinucleated L6 muscle cells with Semliki Forest virus and VSV at high multiplicities showed that the glycoprotein of each virus occupied intracellular domains which were devoid of the other respective glycoprotein. Taken together, these findings indicate that the viral glycoproteins did not range far from their site of synthesis within the ER or other intracellular membrane compartments in these large cells. This result also suggests that relocation of viral RNA synthesis occurred slowly. 相似文献
15.
Macrophages are generated through the differentiation of monocytes in tissues and they have important functions in innate and adaptive immunity. In addition to their roles as phagocytes, macrophages can be further differentiated, in the presence of receptor activator of nuclear factor kappa-B ligand (RANKL) and macrophage colony-stimulating factor (M-CSF), into osteoclasts (multinucleated giant cells that are responsible for bone resorption). In this work, we set out to characterize whether various inflammatory stimuli, known to induce macrophage polarization, can alter the type of multinucleated giant cell obtained from RANKL differentiation. Following a four-day differentiation protocol, along with lipopolysaccharide (LPS)/interferon gamma (IFNγ) as one stimulus, and interleukin-4 (IL-4) as the other, three types of multinucleated cells were generated. Using various microscopy techniques (bright field, epifluorescence and scanning electron), functional assays, and western blotting for osteoclast markers, we found that, as expected, RANKL treatment alone resulted in osteoclasts, whereas the addition of LPS/IFNγ to RANKL pre-treated macrophages generated Langhans-type giant cells, while IL-4 led to giant cells resembling foreign body giant cells with osteoclast-like characteristics. Finally, to gain insight into the modulation of osteoclastogenesis, we characterized the formation and morphology of RANKL and LPS/IFNγ-induced multinucleated giant cells. 相似文献
16.
Suparak S Muangsombut V Riyapa D Stevens JM Stevens MP Lertmemongkolchai G Korbsrisate S 《Microbes and infection / Institut Pasteur》2011,13(12-13):1006-1011
Burkholderia pseudomallei induces the formation of multinucleated giant cells in cell monolayers. After infection of human macrophage-like U937 cells with B. pseudomallei, addition of monoclonal antibodies against integrin-associated protein (CD47), E-selectin (CD62E), a fusion regulatory protein (CD98), and E-cadherin (CD324) suppressed multinucleated giant cells in a concentration-dependent manner while monoclonal antibodies against other surface molecules did not inhibit fusion despite binding to the cell surface. Flow cytometric analysis showed increased expression of CD47 and CD98, but not CD62E and CD324, upon B. pseudomallei infection. Our data suggest the involvement of specific cellular factors in the process of B. pseudomallei-induced fusion. 相似文献
17.
Yoshida K Obata S Ono M Esaki M Maejima T Sawada H 《European journal of cell biology》2007,86(8):461-471
The 5F9A cell, which is a mesenchymal stem cell-like clone established from rat bone marrow substrate adherent cells, can differentiate into adipocytes and osteoblasts in vitro under the appropriate conditions. Multinucleated cells could be also induced by 12-O-tetradecanoylphorbol 13-acetate (TPA) in 5F9A cells. This effect was mediated by protein kinase C. Possible mechanisms of multinucleation by TPA were hypothesized to be either karyokinesis without cytokinesis or cell-cell fusion. By observation using time-lapse phase-contrast microscopy, we determined that the multinucleated cells were generated mainly by karyokinesis without cytokinesis. Cell fusion was studied using time-lapse photography, and confocal laser scanning microscopy using two differentially labeled cells. These techniques demonstrated that multinucleated 5F9A cells could be produced by cell fusion, albeit at a low frequency. We conclude that multinucleated 5F9A cells are formed primarily by karyokinesis without cytokinesis, although some cells are also formed by cell-cell fusion. 相似文献
18.
Summary Mononucleated cells derived from canine bone marrow were maintained in vitro for up to 6 weeks. The culture characteristics and development of these cells were evaluated by histological, ultrastructural and histochemical methods. Within 1 week the cells had fused together to form flattened, multinucleated cells. Further fusing with one another and other mononucleated cells produced large (diameters more than 300 m), multinucleated cells which frequently contained more than 50 nuclei per cell and exhibited ultrastructural and histochemical features that were strikingly similar to those displayed by osteoclasts. The confluent monolayer of mono- and multinucleated cells present at 4 weeks had, by the sixth week, become altered such that fibroblast overgrowth obliterated all other cells. During the development of the culture adipocytes became differentiated from mononuclear cells and frequently were located within spherical multicellular aggregates (spheroids). Functional assessments were employed to investigate whether the multinucleated cells generated in this way, represented osteoclast-like cells, or alternatively, were related to macrophage polykarya as found in foreign body granulomata in vivo. Neither resorption pits on sperm whale dentine slivers (diagnostic of osteoclasts), nor formation of granulomata in vitro, were observed. We believe that the present results indicate that the multinucleated cells generated from canine bone marrow mononuclear precursors in vitro, merit designation as osteoclast-like cells. Definitive characterisation however, must await further functional assessments of hormone responsiveness.Dr. Martin Bird died during the final phase of this work and it is to his memory that this paper is respectfully dedicated 相似文献
19.
Characterization of chimeras between the ecotropic Moloney murine leukemia virus and the amphotropic 4070A envelope proteins. 总被引:4,自引:4,他引:0
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A series of 22 chimeric envelope (env) genes were generated between the ecotropic Moloney murine leukemia virus and the amphotropic 4070A isolate. The chimeric envelopes were expressed within the complete, replication-competent provirus and tested for virus viability by transient expression assays. Eleven of the 22 viruses were viable. Five of these chimeric viruses showed an ecotropic host range, and six exhibited an amphotropic host range and viral interference. The host range determinants map to the first half of the surface (SU) protein. The N-terminal 72 amino acids of 4070A (42 of processed SU) are not required for amphotropic receptor usage. Ecotropic and amphotropic viruses differ in their ability to form large, multinucleated syncytia when cocultured with the rat XC cell line. Ecotropic murine leukemia virus forms large syncytia with XC cells, whereas no syncytia are reported for amphotropic virus. All chimeras which contained the N-terminal half of the ecotropic SU protein, encoding the receptor binding domain, formed the large multinucleated syncytia with XC cells. 相似文献