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1.
Immunogold staining of primary cultures of human brain microvessel endothelial cells demonstrated the presence of Factor VIII-related antigen within cytoplasmic vesicles in close association with the rough endoplasmic reticulum and Golgi apparatus. Immunoperoxidase staining, at the light microscopic level, revealed a similar granular, perinuclear staining. The morphology and location of these vesicular profiles indicate that they are part of the trans-Golgi region where terminal processing and short-term storage of Factor VIII-related antigen takes place. Weibel-Palade bodies, specific storage organelles for von Willebrand factor in large vessel endothelium, were not observed in cerebral microvessel endothelium. The release of Factor VIII-related antigen from the cytoplasmic vesicles was influenced by some of the factors known to stimulate or inhibit the regulated pathway of secretion from Weibel-Palade bodies. Thus, stimulation of endothelial cells with calcium ionophore A23187 resulted in almost complete loss of staining, while addition of EGTA to the culture medium led to slight increase of intracellular pools of Factor VIII-related antigen. Pre-incubation of monolayers with interferon-gamma was associated with significant increase in the number of labeled vesicles, suggesting an additional role of this cytokine in the localized immune reaction within the central nervous system. 相似文献
2.
Electron-microscopic and immunocytochemical analyses of Weibel-Palade bodies in the human umbilical vein during pregnancy 总被引:3,自引:0,他引:3
Summary The present study was done to elucidate the biological significance of the Weibel-Palade body of human umbilical vein endothelial cells. Quantitative determinations of these endothelial-specific granules throughout pregnancy revealed that their numbers and size per cell profile were maintained at low levels from 12 to 19 weeks of gestation; then both rapidly increased from 33 weeks to full term. This increase coincided with the development of the rough endoplasmic reticulum and an increase in the number of endothelial cell pinocytotic vesicles. Light-microscopic peroxidase anti-peroxidase and electron-microscopic protein A-gold techniques provided evidence that factor VIII-related antigen was localized in the Weibel-Palade bodies. Furthermore, in vitro treatment of incubated umbilical vein tissue with compound 48/80, a histamine releaser, induced degranulation of Weibel-Palade bodies from the endothelium. The present study indicates that Weibel-Palade bodies are storage sites of both histamine and factor VIII-related antigen and have an important role in the obliteration of this vessel. 相似文献
3.
Summary Lymphatic endothelial cells isolated from bovine mesenteric lymphatic vessels were cultured and characterized. Lymphatic endothelial
cells grew as a monolayer displaying an elongated morphology in preconfluent primary cultures. When confluent, the cells exhibited
a polygonal morphology to form a “cobblestone” pattern previously described for cultured vascular endothelium. All culture
lymphatic endothelial cells expressed Factor VIII-related antigen and boundUlex europaeus I lectin. Ultrastructurally, cultured lymphatic endothelium was characterized by the presence of Weibel-Palade bodies as
well as the usual cytoplasmic organelles. 相似文献
4.
H Ide Y Minamishima Y Eizuru M Okada K Sakihama T Katsuki 《Microbiology and immunology》1988,32(1):45-55
Human endothelial cells derived from the umbilical vein were transformed with SV40 virions. A cell line subcultured for over 60 serial passages was characterized in comparison with its untransformed counterpart which was culturable for less than five passages. The SV40-transformed human endothelial cells, designated SV-HUVEC, were positive not only for tumor (T) antigen specific to the SV40-transformed cell, but also for two markers of endothelial cells, Factor VIII-related antigen and a receptor for Ulex europaeus agglutinin I. By transformation the growth potential of the human endothelial cells was increased and their serum requirement was decreased. The SV40-transformed endothelial cells were, however, unable to form colonies in soft agar or to form tumors in athymic nude mice, although a small nodule was produced at the site of inoculation. Subcultivation of these cells up to the 62nd passage eventually resulted in crisis and loss of further cell division. Thus, the human endothelial cells were transformed by SV40 while retaining certain normal functions but without showing tumorigenicity. 相似文献
5.
Isolation of a new serotype of simian acquired immune deficiency syndrome type D retrovirus from Celebes black macaques (Macaca nigra) with immune deficiency and retroperitoneal fibromatosis. 总被引:9,自引:6,他引:3
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P A Marx M L Bryant K G Osborn D H Maul N W Lerche L J Lowenstine J D Kluge C P Zaiss R V Henrickson S M Shiigi et al. 《Journal of virology》1985,56(2):571-578
A new serotype of simian acquired immune deficiency syndrome (SAIDS) retrovirus (type 2) belonging to the D genus of retroviruses is associated with a SAIDS occurring spontaneously in a colony of Celebes macaques (Macaca nigra) and rhesus macaques (Macaca mulatta) at the Oregon Regional Primate Research Center. This syndrome resembles SAIDS in M. mulatta at the California Primate Research Center, which is associated with a similar type D retrovirus (type 1). However, at the Oregon Center, SAIDS is distinguished by the occurrence of retroperitoneal fibromatosis in some of the affected monkeys. Type 2 virus was isolated from seven of seven macaques with SAIDS, retroperitoneal fibromatosis, or both and from one of six healthy macaques. The new strain is closely related to SAIDS retrovirus type 1 and Mason-Pfizer monkey virus but can be distinguished by competitive radioimmunoassay for minor core (p10) antigen and by genomic restriction endonuclease cleavage patterns. Neutralization tests indicate that type 1 and type 2 SAIDS retroviruses are distinct serotypes. Therefore, separate vaccines may be necessary to control these infections in colonies of captive macaques. 相似文献
6.
Cultured endothelial cells derived from the human iliac arteries 总被引:1,自引:0,他引:1
M. K. Glassberg M. M. Bern S. R. Coughlin C. C. Haudenschild L. W. Hoyer H. N. Antoniades B. R. Zetter 《In vitro cellular & developmental biology. Plant》1982,18(10):859-866
Summary Cells derived from the endothelium of human iliac arteries were cultured in vivo. The cells were isolated, grown, and subcultured
in HEPES buffered Medium 199 supplemented with 20% heat inactivated human whole blood serum, human alpha-thrombin, and commercial
endothelial cell growth supplement derived from bovine brain. The cells were viable in culture for 8 to 10 passages at a split
ratio of 1:3. After the 10th passage, the cells began to enlarge and their growth rate was reduced. No cultures were viable
after the 12th passage. The cells were determined to be of endothelial origin by their morphology at confluence; their ultrastructural
characteristics, including the presence of Weibel-Palade bodies; the production and release of factor VIII-related antigen;
and by their maintenance of a surface that prevented platelet attachment. The cultured arterial endothelial cells released
prostacyclin in response to challenge with thrombin and protamine sulfate but not in response to bradykinin or the platelet-derived
growth factor. Although the cultures described in this report were derived from patients with varying degrees of atherosclerotic
disease, there were no significant differences in morphological or physiological parameters among these cultures or in comparison
with commonly studied cells derived from human umbilical veins.
The above work was supported by Grant CA28540 from the National Institutes of Health and by a grant from The Council for Tobacco
Research, USA. 相似文献
7.
B. V. Sapatino C. J. R. Welsh C. A. Smith B. F. Bebo D. S. Linthicum 《In vitro cellular & developmental biology. Animal》1993,29(12):923-928
Summary This communication describes a relatively novel cell culture technique for the isolation of cerebrovascular endothelial cells
from three strains of inbred mice. Cerebrovascular endothelial cells were identified by their morphology, the presence of
Factor VIII-related antigen and angiotensin-converting enzyme, and the uptake of acetylated low-density lipoprotein. Cloned
cerebrovascular endothelial cells were found to maintain their differentiated state and diploid genotype through 15 serial
passages. The morphology and growth characteristics of these cells were found to be altered when cultured on different extracellular
matrices. The isolation and cloning methods described are simple and highly reproducible. 相似文献
8.
Isolation and long-term serial cultivation of endothelial cells from the microvessels of the adult human dermis 总被引:1,自引:0,他引:1
P. M. Davison K. Bensch M. A. Karasek 《In vitro cellular & developmental biology. Plant》1983,19(12):937-945
Sumamry A method to isolate and maintain microvascular endothelial cells from the cutaneous vessels of adult human skin in long-term
culture has been developed. Endothelial cells lining the microvessels of the papillary dermis are released from surrounding
tissue during a brief trypsin incubation (0.3% trypsin, 1% EDTA). Cells are plated onto a fibronectin substrate and maintained
in Leibovitz (L15) culture medium containing pooled human serum (50%) and antibiotics. Proliferation is dependent upon the
presence of several additional growth factors, cholera enterotoxin (1×10−9
M), isobutyl methylxanthine (3.3×10−5
M), and medium conditioned by explant culture of the mouse EHS sarcoma. Using this supplemented medium, cells proliferate readily
and can be cultivated serially for more than 6 passages (3 months in vitro). These cells retain their characteristic endothelial
cell morphology, stain positively for Factor VIII antigen, and contain Weibel-Palade bodies.
This research was supported by grant AG 01312 from the U.S. Public Health Service, Washington, D.C. 相似文献
9.
Dennis Way Mary Hendrix Marlys Witte Charles Witte Ray Nagle John Davis 《In vitro cellular & developmental biology. Plant》1987,23(9):647-652
Summary An endothelial cell line derived from a massive recurrent chyle-containing retroperitoneal lymphangioma was isolated in monolayer
culture. Scanning and transmission electron microscopy and immunohistochemistry confirmed a close resemblance to blood vascular
endothelium with typical cobblestone morphology, positive immunofluorescence staining for endothelial marker Factor VIII-associated
antigen and fibronectin, and prominent Weibel-Palade bodies. The endothelial cells also exhibited other ultrastructural features
characteristic of lymphatic endothelium, including sparse microvillous surface projections, overlapping intercellular junctions,
and abundant intermediate filaments. This endothelial cell line represents a new source of proliferating lymphatic endothelium
for future study, including structural and functional comparison to blood vascular endothelium.
Supported in part by Arizona Disease Control Research Commission contracts 8277-000000-1-1-AT-6625 and ZB-7492. Presented
in part at the 10th International Congress of Lymphology in Adelaide, Australia, August 1985. 相似文献
10.
Isolation and long-term serial cultivation of endothelial cells from the microvessels of the adult human dermis 总被引:2,自引:0,他引:2
A method to isolate and maintain microvascular endothelial cells from the cutaneous vessels of adult human skin in long-term culture has been developed. Endothelial cells lining the microvessels of the papillary dermis are released from surrounding tissue during a brief trypsin incubation (0.3% trypsin, 1% EDTA). Cells are plated onto a fibronectin substrate and maintained in Leibovitz (L15) culture medium containing pooled human serum (50%) and antibiotics. Proliferation is dependent upon the presence of several additional growth factors, cholera enterotoxin (1 X 10(-9) M), isobutyl methylxanthine (3.3 X 10(-5) M), and medium conditioned by explant culture of the mouse EHS sarcoma. Using this supplemented medium, cells proliferate readily and can be cultivated serially for more than 6 passages (3 months in vitro). These cells retain their characteristic endothelial cell morphology, stain positively for Factor VIII antigen, and contain Weibel-Palade bodies. 相似文献
11.
Kimiko Takahashi Yoshio Sawasaki Jun-Ichi Hata Kiyoshi Mukai Tamotsu Goto 《In vitro cellular & developmental biology. Plant》1990,26(3):265-274
Summary A new cell line from the human umbilical vein has been established and maintained for more than 5 yr (180 generations; 900
population doublings). This strain, designated ECV304, is characterized by a cobblestone monolayer growth pattern, high proliferative
potential without any specific growth factor requirement, and anchorage dependency with contact inhibition. Karyotype analysis
of this cell line reveals it to be of human chromosomal constitution with a high trisomic karyotype (mode 80). Ultrastructurally,
endothelium-specific Weibel-Palade bodies were identified. Although one of the endothelial cell markers, Factor VIII-related
antigen (VIIIR:Ag) was negative in this cell line, immunocytochemical staining for the lectin Ulex europaeus I (UEA-I), and
PHM5 (anti-human endothelium as well as glomerular epithelium monoclonal antibody) was positive, and angiotensin-converting
enzyme (ACE) activity was also demonstrated. In addition, ECV304 displayed negativity for alkaline and acid phosphatase and
for the epithelial marker keratin. All of these findings suggest that ECV304 cells originated from umbilical vein endothelial
cells by spontaneous transformation. Ultrastructurally, no viruslike particles have been detected intracellularly. Nude mouse
tumorigenicity and rabbit cornea tests were both positive. This is a report on a novel case of phenotypic alteration of normal
venous endothelial cells of human origin in vitro, and generation of a transformant with indefinite life spans. This line
may be useful in studies of some physiologically active factors available for medical use. 相似文献
12.
Canine teeth were extracted from seven adult male Japanese monkeys. Observations over the next four years led to the conclusion that canines are not essential for either the attainment or maintenance of high rank, but that they may play an important part in the self-defense of low-ranking males.Publication No.501 of the Oregon Regional Primate Research Center supported by NIH Grant RR 00163. 相似文献
13.
Twinning pedigrees for Lemur catta and Galago crassicaudatus argentatus colonies at the Oregon Regional Primate Research Center have been constructed. Unlike-sexed twin pairs were more numerous than like-sexed twin pairs in both species. Evidence is presented which supports the hypothesis that twinning in prosimians, as in man, is genetically controlled. 相似文献
14.
Boro Dropulić Colin L. Masters 《In vitro cellular & developmental biology. Plant》1987,23(11):775-781
Summary The isolation and culture of cell lines from mouse brain capillary endothelium (MBE) is described. Cells migrating from collagenase-treated
capillary fragments proliferated rapidly in the 1st wk of culture forming large epithelioid cobblestonelike colonies. The
cells showed only marginal proliferation after 2 to 3 wk in culture, until peripheral cells migrated away from the colony
which exhibited a marked degree of proliferation. These cells were trypsinized and subcultured to confluence. The cells can
be maintained for well over 40 passages and seem to retain their endothelial morphology. The endothelial origin of these cells
was demonstrated by positive immunoperoxidase reactivity with Factor VIII-related antigen, specific binding ofBandeiraea simplicifolia lectin and γ-glutamyl transpeptidase activity. Electron microscopic examination of the MBE cells showed junctional complexes
including intermediate junctions, but no tight junctions. The overall ultrastructure indicates that a degree of dedifferentiation
has occurred, the cells ultrastructurally resembling immature endothelium. An earlier investigation of cultured mouse brain
endothelial cells reported a cell line that had lost many functional and structural characteristics. Our study demonstrates,
as the previous one, that a certain degree of dedifferentiation needs to occur if MBE cells are to be maintained for long-term
culture. However, the degree of dedifferentiation seems to be variable, depending in part on the culture conditions employed.
This work was supported in part by grants from the National Health and Medical Research Council of Australia and the Telethon
and Royal Perth Hospital Research Foundations. 相似文献
15.
Wolf H. Fahrenbach Ph.D. 《Cell and tissue research》1981,216(3):655-659
Summary Efferent fibers to the compound eye of the horseshoe crab, Limulus polyphemus, not only innervate the various pigment cells, but also invade the eccentric cell dendrite and the retinula cells. This finding provides a structural basis for the coupling of circadian rhythm between the efferents and the receptor cells.This short communication is Publication No. 1130 of the Oregon Regional Primate Research Center. The research was supported by Grants RR-00163 and EY-00392 from the National Institutes of Health 相似文献
16.
The social structure of the Oregon troop of Japanese macaques 总被引:1,自引:1,他引:0
Intensive observations were made of a troop ofM. fuscata which was kept in a large outdoor corral at the Oregon Regional Primate Research Center during the summer and fall of 1967. The intent of the observations was to determine whether the troop's normal patterns of social behavior and social structure seemed to be affected by confinement in the corral. Substantial deviations were noted between the spatial structure of the Oregon troop and the central-peripheral structure typical of free-ranging troops in Japan. There was also some indication of heightened aggressiveness within the corral but other aspects of social behavior seemed unaltered. 相似文献
17.
M K Glassberg M M Bern S R Coughlin C C Haudenschild L W Hoyer H N Antoniades B R Zetter 《In vitro》1982,18(10):859-866
Cells derived from the endothelium of human iliac arteries were cultured in vivo. The cells were isolated, grown, and subcultured in HEPES buffered Medium 199 supplemented with 20% heat inactivated human whole blood serum, human alpha-thrombin, and commercial endothelial cell growth supplement derived from bovine brain. The cells were viable in culture for 8 to 10 passages at a split ratio of 1:3. After the 10th passage, the cells began to enlarge and their growth rate was reduced. No cultures were viable after the 12th passage. The cells were determined to be of endothelial origin by their morphology at confluence; their ultrastructural characteristics, including the presence of Weibel-Palade bodies; the production and release of factor VIII-related antigen; and by their maintenance of a surface that prevented platelet attachment. The cultured arterial endothelial cells released prostacyclin in response to challenge with thrombin and protamine sulfate but not in response to bradykinin or the platelet-derived growth factor. Although the cultures described in this report were derived from patients with varying degrees of atherosclerotic disease, there were no significant differences in morphological or physiological parameters among these cultures or in comparison with commonly studied cells derived from human umbilical veins. 相似文献
18.
Nancy J. Alexander 《Cell and tissue research》1973,136(2):177-182
Summary Epididymal biopsies from rats that had undergone unilateral or bilateral vasectomies from one to eight months previously were compared with biopsies from their contralateral side or from normal controls to ascertain what ultrastructural changes had occurred. After vasectomy, spermatozoa appeared to dissolve in the lumen of the caput epididymidis and to be absorbed by the principal cells. About 5 weeks after vasectomy, numerous lamellar accumulations became apparent in the supernuclear region. Their resemblance to lysosomes or residual bodies was confirmed by an acid phosphatase reaction. After 10 weeks, similar lamellar and polymorphic accumulations on the contralateral side of animals with unilateral vasectomies indicated that resorption had also increased on the unligated side.Publication No. 627 of the Oregon Regional Primate Research Center. This study was supported by NIH Grants No. RR-00163 and HD-05969.The author wishes to thank Ms. J. Hren for her excellent technical assistance. 相似文献
19.
Originally described in vascular endothelial cells, Weibel-Palade bodies were considered as specific of this cellular type, as they have never been reported elsewhere. Weibel-Palade bodies serve as storage granules for von Willebrand factor which is stored in microtubular form. Besides endothelial cells von Willebrand factor is also synthetized by bone marrow megakaryocytes. Von Willebrand factor has been located in alpha-granules of megakaryocytes and blood platelets. We describe true Weibel-Palade bodies in pig megakaryocytes, and also alpha-granules which look like an evolutionary form of Weibel-Palade bodies. Von Willebrand Factor is most likely stored in microtubular form in these two types of structure. This is supported by the absence of microtubules in these granules in cells obtained from pigs homozygous for the von Willebrand disease (lacking totally this protein). 相似文献
20.
Starch gel electrophoresis of erythrocytes from 1812 Macaca mulatta has unequivocally demonstrated that the 6-phosphogluconate dehydrogenase (6PGD) isozymes are controlled by two autosomal codominant alleles. Limited data on erythrocytes from 89 Macaca speciosa were also consistent with autosomal codominance.This work was supported in part by NIH Grants HD 07835 (WHS) and RR-00167 (Wisconsin Regional Primate Research Center) and by the Research Committee of the UW Graduate School (Project No. 170207).Paper No. 2146 of the Laboratory of Genetics, and Publication No. 16-045 of The Wisconsin Regional Primate Research Center. 相似文献