Heparin Enhances Endothelial Cell von Willebrand Factor Content by Growth Factor Dependent Mechanisms

Tannenbaum, Susan; Chao, Elizabeth S.; Gralnick, Harvey R.

doi:10.1055/s-0038-1648956

Cited by 11 publications

(9 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The vWF, which was found only in endothelial cells, platelets and megakaryocytes (Tannenbaum et al., 1994), is considered the most reliable marker for endothelial cells in culture (Folkman and Klagsbrun, 1987; Ulger et al., 1997). For some time, the terminology applied to factor VIII and vWF was confusing.…”

Section: Discussionmentioning

confidence: 99%

“…The terminology was complicated by the fact that when the complex was initially isolated, the activity measured was that of factor VIII (procoagulant activity), but the protein detected was vWF (Ruggeri, 1991). vWF is stored as extremely high molecular weight multimers in Weibel–Palade bodies which are endothelium‐specific (Meyer et al., 1991; Tannenbaum et al., 1994). Immunochemical staining for vWF showed that it reacts with endothelial cells from several mammalian species (Gentile et al., 1998; Zhang et al., 1998; De Meyer et al., 1999; Hanke et al., 1999; Lamszus et al., 1999; Vailhé et al., 1999).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Labelling of Rat Endothelial Cells with Antibodies to vWF, RECA‐1, PECAM‐1, ICAM‐1, OX‐43 and ZO‐1

Ülger

Karabulut

Pratten

2002

Anat Histol Embryol

View full text Add to dashboard Cite

Labelling with endothelium specific monoclonal antibodies, von Willebrand Factor (vWF), rat endothelial cell antigen-1 (RECA-1), platelet-endothelial cell adhesion molecule-1 (PECAM-1), intercellular adhesion molecule-1 (ICAM-1), OX-43 and zonula occludentes-1 (ZO-1), was investigated in cryostat sections of vessels from rats of different ages using a confocal microscope. The results showed that labelling of the vWF was positive in endothelial cells from adult, fetal and different ages of embryonic rat. Labelling with RECA-1 was weakly positive in adult rat aorta and lung endothelial cells but not in embryonic yolk sac endothelial cells. Labelling using PECAM-1, ICAM-1 and OX-43 was negative in both adult and embryonic endothelial cells. ZO-1 showed positive but very weak reactivity in embryonic yolk sac endothelial cells. The expression of vWF on vessels from adult and 19.5-day fetal tissues was strongly positive. However, the expression of vWF in embryonic endothelial cells was dependent on the gestational age. While the 11.5-day yolk sac vessels stained weakly, staining gradually increased in 13.5-, 15.5- and 17.5-day-old yolk sac vessels. The results suggest that vWF is a reliable endothelial cell marker in rat vascular endothelial cells, including both fetal and embryonic stages.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Labelling of Rat Endothelial Cells with Antibodies to vWF, RECA‐1, PECAM‐1, ICAM‐1, OX‐43 and ZO‐1

Ülger

Karabulut

Pratten

2002

Anat Histol Embryol

View full text Add to dashboard Cite

show abstract

“…Therefore, a general endothelial cell marker has not been found yet. Immunochemical staining for vWF, which was found only in endothelial cells, platelets, and megakaryocytes (26), is considered the most reliable marker for endothelial cells in culture (27). vWF reacts specifically with the cytoplasm of human endothelial cells of normal, reactive and neoplastic blood and lymphatic vessels.…”

Section: Discussionmentioning

confidence: 99%

Isolation and culture of endothelial cells from embryonic rat yolk sac

Ülger

Karabulut

Pratten

2017

The EuroBiotech Journal

View full text Add to dashboard Cite

Yolk sac blood islands are the first morphologic evidence of hematopoietic development during mammalian embryogenesis, and visseral yolk sac mesoderm gives rise to the first embryonic blood cells within a rich endothelial network. Present study reports the isolation and culture of endothelial cells from 11.5 days old embryonic rat yolk sac. The embryos were dissected from 11.5 days pregnant Wistar rat (Rattus norvegicus) and the external yolk sac membrane and embryos were removed under aseptic condition. After washing three times with Calcium-Magnesium free Hank's balanced salt solution (CMF-HBSS), the tissue was minced, and fragments were incubated in CMF-HBSS containing 2mg/ml Trypsin, 100mg/ml collagenase I and 40mg/ml DNAse at 37°C until the tissue was completely dispersed. The digestion effect was then neutralized by fetal bovine serum at 1:3 (v/v). The cell suspension was centrifuged at 1000 rpm for 10 min., the supernatants were discarded and the cell pellets resuspended in Dulbecco modified Eagle medium containing 15% fetal bovine serum, 1.25mg/ml amphotericin B, 25mg/ml gentamycin sulphate and 100mg/ml endothelial cell growth supplement. The resuspended cells were plated in two diverse 25cm 2 culture flasks for overnight differential adherence at 37°C. The non-adherent cells were removed by gentle aspiration and adherent cells refed with fresh medium. The cells were transferred using 1ml of 0.2% Trypsin when cultures reached near-confluence. The cultured yolk sac endothelial cells had characteristic cobblestone appearence and positive immunofluorescent staining for von Willebrand Factor (vWF). Weibel-Palade bodies, the major ultrastructural marker for endothelium, were also detected in cultured cells by electron microscopy.

show abstract

“…The rapid PAI-1 mRNA decrease initially observed with low-PAI-1 mRNA decrease might be related to an inhibitory effect of heparin on enzymes present in dose heparin as well as the significant delay and long-lasting reduction observed with higher UFH the RT-PCR [28,29] can be ruled out since G3PDH amplification was not affected. doses can be explained by the different modulation of some endothelial cell properties in the presence Studies evaluating the changes in vascular endothelium induced by glycosaminoglycans have foof heparin: a transient effect, probably related to the presence of heparin molecules bound at the cused primarily on the modulation of the procoagulant properties of stimulated endothelial cultures membrane surface, and a delayed one mediated in the presence of heparin [30][31][32]. Our results also of PAI-1, which could explain some of its profibrinolytic properties observed in vivo [18,19].…”

Section: Effect Of Ufh Andmentioning

confidence: 99%

Evidence that Heparin but Not Hirudin Reduces PAI-1 Expression in Cultured Human Endothelial Cells

Orbe¹,

Montes²,

Zabalegui³

et al. 1999

Thrombosis Research

View full text Add to dashboard Cite

in the regulation of fibrinolysis through the 10 IU/ml) and hirudin (20 and 100 TIU/ml). Samples synthesis and secretion of plasminogen actiwere obtained before and 2, 6, and 24 hours after vators (tissue plasminogen activator [t-PA] and urostimulation. mRNA analysis was conducted by rekinase plasminogen activator [u-PA]) and plasminoverse transcription followed by polymerase chain gen activator inhibitor-1 (PAI-1) [1]. Several studies reaction, and PAI-1 antigen was determined by suggest that PAI-1, the main plasminogen activator ELISA. Addition of UFH (10 IU/ml) to HUVEC inhibitor found in plasma, plays an important pathoresulted in a decrease of PAI-1 mRNA at 6 hours physiological role. Raised plasma levels have been (40% reduction) and 24 hours (60% reduction) and associated with thrombosis [2-4], while defective PAI-1 antigen. Hirudin, however, did not modify PAI-1 is associated with a bleeding tendency [5]. significantly the PAI-1 mRNA nor the inhibitor se-PAI-1 is a 50-kD glycoprotein containing 379 cretion. The addition of UFH (10 or 100 IU/ml) aminoacids and 3 N-glycosylation sites, which exto endotoxin-stimulated HUVEC also reduced the ists in different conformations in vivo. Endothelial increased PAI-1 mRNA and antigen secretion cells synthesize an active form that spontaneously (45%), whereas no effect could be observed with converts to the latent or inactive form due to conhirudin. Our results suggest that UFH, but not formational changes of protein tertiary structure hirudin, by reducing the endothelial expression of [6,7]. The PAI-1 gene is located in chromosome 7, and it has 12.2 kb organized in nine exons separated by eight introns [8,9]. The 3Ј untranslated region

show abstract

Heparin Enhances Endothelial Cell von Willebrand Factor Content by Growth Factor Dependent Mechanisms

Cited by 11 publications

References 37 publications

Labelling of Rat Endothelial Cells with Antibodies to vWF, RECA‐1, PECAM‐1, ICAM‐1, OX‐43 and ZO‐1

Labelling of Rat Endothelial Cells with Antibodies to vWF, RECA‐1, PECAM‐1, ICAM‐1, OX‐43 and ZO‐1

Isolation and culture of endothelial cells from embryonic rat yolk sac

Evidence that Heparin but Not Hirudin Reduces PAI-1 Expression in Cultured Human Endothelial Cells

Contact Info

Product

Resources

About