scholarly journals Endothelial cell mitogens derived from retina and hypothalamus: biochemical and biological similarities.

1984 ◽  
Vol 99 (4) ◽  
pp. 1545-1549 ◽  
Author(s):  
P A D'Amore ◽  
M Klagsbrun
Keyword(s):  
Endothelial Cell ◽  
Growth Factor ◽  
Affinity Chromatography ◽  
High Performance ◽  
Biological Activities ◽  
Umbilical Vein ◽  
Size Exclusion ◽  
3T3 Cells ◽  
Two Factors ◽  
Heparin Affinity

Bovine retina and hypothalamus contain anionic endothelial cell mitogens that display unusual affinities for the negatively charged glycosaminoglycan heparin. Both growth factor activities are acidic polypeptides (pl's of 5.0) as determined by isoelectric focusing and DEAE-affinity chromatography. In spite of their anionic nature, the factors bound to heparin-Sepharose columns with high affinity and could be eluted only at high salt concentrations (0.9-1.1 M NaCl). The affinity of the retina-derived growth factor (RDGF) for heparin permitted a 15,000-fold purification of the mitogen in two steps: heparin-affinity chromatography and size exclusion high-performance liquid chromatography. RDGF and the anionic hypothalamus-derived factor (aHDGF) exhibit three major biochemical similarities including isoelectric point, (pl's of 5.0), heparin affinity (elution at 0.9-1.1 M NaCl) and molecular weight (18,000). Additionally, the two factors display similar biological activities, stimulating the proliferation of capillary and human umbilical vein endothelial and 3T3 cells but not vascular smooth muscle cells. We suggest that RDGF and aHDGF are related if not identical growth factor molecules.

Characterization of heparin-binding endothelial mitogen(s) produced by the ovine endometrium during early pregnancy

1998 ◽  
Vol 76 (1) ◽  
pp. 89-96 ◽  
Author(s):  
Jing Zheng ◽  
Dale A Redmer ◽  
S Derek Killilea ◽  
Lawrence P Reynolds
Keyword(s):  
Growth Factor ◽  
Affinity Chromatography ◽  
Early Pregnancy ◽  
Molecular Size ◽  
Immunoblot Analysis ◽  
Binding Activity ◽  
Mitogenic Activity ◽  
Heparin Binding ◽  
3T3 Cells ◽  
Heparin Affinity

To characterize mitogenic factors produced by ovine endometrium during early pregnancy, endometrial explant-conditioned media (ECM) were obtained from ewes on day 12, 18, 24, or 30 after mating. These ECM contained mitogenic activity for both endothelial and 3T3 cells across all days. The endothelial mitogenic activity was greatest on day 24, whereas mitogenic activity for 3T3 cells did not differ across days. By ultrafiltration, ion exchange, and heparin-affinity chromatography, the endothelial mitogenic activity was found to have a molecular mass greater than 100 kDa, to be anionic, and to be heparin binding, respectively. Three peaks of endothelial mitogenic activity were recovered from heparin-affinity chromatography. The major peak, H3, was mitogenic for endothelial but not for 3T3 cells. H3 was further purified, and the single peak of heparin-binding activity, designated H3b, represented a 681-fold purification of endothelial mitogenic activity from endometrial ECM. H3 and H3b were heat labile and trypsin sensitive, and their biological activity was heparin enhanced. The majority of the endothelial mitogenic activity was immunoneutralized by antibodies against acidic and basic FGF. Nevertheless, we were unable to detect bFGF in H3 or H3b by Western immunoblot analysis. Thus, in this study we have extended our previous observations and demonstrated that (i) during early pregnancy the ovine endometrium produces mitogenic activity for both endothelial and 3T3 cells, (ii) the endothelial mitogenic activity is greatest on day 24 after mating, which corresponds with the onset of endometrial vascular growth, and (iii ) the major endothelial mitogen has a high affinity for heparin, and although it is immunologically related to FGF, it differs from known FGF in its apparent molecular size and biological activities.Key words: heparin-binding growth factor, fibroblast growth factor, uterus, early pregnancy, ewe.

VEGF-C mediates cyclic pressure-induced endothelial cell proliferation

2002 ◽  
Vol 11 (3) ◽  
pp. 245-251 ◽  
Author(s):  
Hainsworth Y. Shin ◽  
Michael L. Smith ◽  
Karen J. Toy ◽  
P. Mickey Williams ◽  
Rena Bizios ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Growth Factor ◽  
Gene Transcription ◽  
Molecular Mechanisms ◽  
Transforming Growth Factor ◽  
Umbilical Vein ◽  
Culture Conditions ◽  
Endothelial Cell Proliferation ◽  
Cyclic Pressure ◽  
Cell Functions

Mechanical forces modulate endothelial cell functions through several mechanisms including regulation of gene transcription. In the present study, gene transcription by human umbilical vein endothelial cells (HUVEC) either maintained under control pressure (that is, standard cell culture conditions equivalent to 0.15 mmHg sustained hydrostatic pressure) or exposed to 60/20 mmHg sinusoidal pressures at 1 Hz were compared using Affymetrix GeneChip microarrays to identify cellular/molecular mechanisms associated with endothelial cell responses to cyclic pressure. Cyclic pressure selectively affected transcription of 14 genes that included a set of mechanosensitive proteins involved in hemostasis (tissue plasminogen activator), cell adhesion (integrin-α2), and cell signaling (Rho B, cytosolic phospholipase A2), as well as a unique subset of cyclic pressure-sensitive genes such as vascular endothelial growth factor (VEGF)-C and transforming growth factor (TGF)-β2. The present study also provided first evidence that VEGF-C, the most highly induced gene under 60/20 mmHg, mediated HUVEC proliferation in response to this cyclic pressure. Cyclic pressure is, therefore, a mechanical force that modulates endothelial cell functions (such as proliferation) by activating a specific transcriptional program.

MODULATION of PROSTACYCLIN PRODUCTION BY HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS WITH ECGF/HEPARIN MEDIUM : ROLE OF CELLULAR DENSITY AT CONFLUENCE

1987 ◽  
Author(s):  
O BOUTHERIN-FALSON ◽  
N BLAES
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Growth Factor ◽  
Umbilical Vein ◽  
Calf Serum ◽  
Lower Amount ◽  
Endothelial Cell Proliferation ◽  
Human Umbilical Vein ◽  
Umbilical Vein Endothelial Cells

Prostacyclin (PGI2) is a major product of arachidonic acid metabolism in vascular endothelial cells. In addition to the role of exogenous agents, its production could be modulated by culture conditions : proliferative state, medium renewal, subcultivation... The use of endothelial cell growth factor (ECGF) associated with heparin has been shown to improve human endothelial cell proliferation. Here we report that human umbilical vein endothelial cells (HUVEC) grown in that medium produce less prostacyclin than without growth factor.HUVEC were cultured in RPMI-199 1:1 + 20% fetal calf serum, added or not with ECGF (Bovine hypothalamus extract BTI Cambridge, 24 ug/ml) and heparin (from porcine intestinal mucosa, Signa, 90 ug/ml). After 4 days in culture, medium was removed and replaced by Tyrode Hepes buffer and basal production was measured after 20 min. Cells were then submitted to 5 min thrombin to assess PGI2 production in stimulated conditions. PGI2 production was estimated by specific radioimmunoassay for 6 keto PGFjalpha. For each point, cell number in the culture was counted after Trypsin EDTA treatment. In the present study, cells grown in ECGF-heparin medium produce lower amount of PGI2, compared to heparin or control medium. This result was observed in both basal and stimulated conditions. For each medium (ECGF-heparin, heparin, control), correlations between PGI2 production per cell and log cell density were shown to be significantly negative.These observations suggest that ECGF effect on PGI2 production could be a consequence of its growth factor activity, notably by the fact that it leads to an endothelial monolayer made of more numerous cells. Since it is now suggested by a number of clinical observations that PGI2 is rather produced in pathological conditions, culture models showing a weak production of PGI2 appear in that connection doser to the physiological conditions.

scholarly journals Dynamic multi-site phosphorylation by Fyn and Abl drives the interaction between CRKL and the novel scaffolding receptors DCBLD1 and DCBLD2

2017 ◽  
Vol 474 (23) ◽  
pp. 3963-3984 ◽  
Author(s):  
Anna M. Schmoker ◽  
Jaye L. Weinert ◽  
Kyle J. Kellett ◽  
Hannah E. Johnson ◽  
Ryan M. Joy ◽  
...  
Keyword(s):  
Growth Factor ◽  
High Performance ◽  
Biological Activities ◽  
Family Members ◽  
Growth Factor Receptor ◽  
Sh2 Domain ◽  
Src Homology 2 ◽  
Negative Role ◽  
Src Homology ◽  
Epidermal Growth

Discoidin, CUB, and LCCL domain containing 2 (DCBLD2) is a neuropilin-like transmembrane scaffolding receptor with known and anticipated roles in vascular remodeling and neuronal positioning. DCBLD2 is also up-regulated in several cancers and can drive glioblastomas downstream of activated epidermal growth factor receptor. While a few studies have shown either a positive or negative role for DCBLD2 in regulating growth factor receptor signaling, little is known about the conserved signaling features of DCBLD family members that drive their molecular activities. We previously identified DCBLD2 tyrosine phosphorylation sites in intracellular YxxP motifs that are required for the phosphorylation-dependent binding of the signaling adaptors CRK and CRKL (CT10 regulator of kinase and CRK-like). These intracellular YxxP motifs are highly conserved across vertebrates and between DCBLD family members. Here, we demonstrate that, as for DCBLD2, DCBLD1 YxxP motifs are required for CRKL–SH2 (Src homology 2) binding. We report that Src family kinases (SFKs) and Abl differentially promote the interaction between the CRKL–SH2 domain and DCBLD1 and DCBLD2, and while SFKs and Abl each promote DCBLD1 and DCBLD2 binding to the CRKL–SH2 domain, the effect of Abl is more pronounced for DCBLD1. Using high-performance liquid chromatography coupled with tandem mass spectrometry, we quantified phosphorylation at several YxxP sites in DCBLD1 and DCBLD2, mapping site-specific preferences for SFKs and Abl. Together, these data provide a platform to decipher the signaling mechanisms by which these novel receptors drive their biological activities.

Influence of Umbilical Cord Blood-Derived Endothelial Precursor Cells on a Human Umbilical Vein Endothelial Cell Co-Culture In Vitro Angiogenesis Assay.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4218-4218
Author(s):  
Nicholas J. Greco ◽  
Brandon Eilertson ◽  
Jason J. Banks ◽  
Paul Scheid ◽  
Marcie Finney ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Growth Factor ◽  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Umbilical Vein ◽  
Tubule Formation ◽  
Human Umbilical Vein ◽  
In Vitro Angiogenesis

Abstract To assess in vitro angiogenesis, cellular co-culture assays have been utilized to study adherence, spreading, differentiation and proliferation, and migration of endothelial cells. Formation of tubule or capillary-like networks is influenced by the presence of vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF) but other factors provided by cell sources and/or direct contact with multiple cell types may facilitate this formation. The hypothesis of this study is that umbilical cord blood (UCB)-derived endothelial precursor cells (EPCs) may influence the formation of human umbilical vein endothelial cell (HUVEC) tubule structures during angiogenesis. Methods: UCB-derived EPCs were isolated from CD133negative cells after a 7-day culture on human fibronectin in EGM-2 media. Tubule formation was evaluated (passage 1–2, 20 x 103 or 2 x 103 cells) by adding HUVECs without or with EPCs to cultures of human bone marrow-derived mesenchymal stromal cells (MSCs) under normoxic (20%) conditions (37°C, 5% CO2, containing VEGF, epidermal growth factor, FGF, insulin-like growth factor, heparin, hydrocortisone, and ascorbic acid in EGM-2 medium) for a 2-week period. HUVECs were added to cultures without or with labeling with Vybrant® CM-DiI which allows the temporal observation of tubule formation progress and cellular incorporation. Final tubule formation was confirmed using a primary anti-CD31 (PECAM) antibody followed by a FITC-conjugated secondary antibody for signal amplification. Results: After 2–4 days, linear aggregates of labeled HUVECs (2-D arrangement) were observed. After 14 days, there was remodeling of HUVECs into the development of a 3D network of linear and branched tubule structures. EPCs facilitated the formation of tubules affecting both the extent of tubule formation and also enhanced proliferation of HUVEC cells. A minority (< 5%) of EPCs were incorporated into developing tubules (estimated using CM-Dil-labeled EPCs). To quantify tubule formation, digital pictures of representative areas of culture wells (2–4/well) were acquired. Using Image Pro Plus software, tubules were quantified using multi-parameter analysis with respect to length, area, and perimeter. The presence of EPCs (equal to the number of added HUVECs) significantly enhanced all parameters. In comparison to control samples, the presence of EPCs increased the area, perimeter and size by 15.2-fold, 3.4-fold, and 3.2-fold, respectively. Confocal microscopy revealed that the co-cultures formed anatamoses, indicating the formation of a connected network. Conclusions: Taken together, these results suggest that the presence of cord blood-derived EPCs facilitate tubule formation and development via a heterotypic cell-cell interaction without integrating into the angiogenic structures. Further studies will evaluate the secretion of cytokines and growth factors.

scholarly journals Dissociation of the chemotactic and mitogenic activities of platelet-derived growth factor by human neutrophil elastase.

1985 ◽  
Vol 100 (2) ◽  
pp. 351-356 ◽  
Author(s):  
R M Senior ◽  
J S Huang ◽  
G L Griffin ◽  
T F Deuel
Keyword(s):  
Growth Factor ◽  
Neutrophil Elastase ◽  
Biological Activities ◽  
Chemotactic Activity ◽  
Mitogenic Activity ◽  
Cathepsin G ◽  
Platelet Derived Growth Factor ◽  
Human Neutrophil Elastase ◽  
3T3 Cells

Because platelet-derived growth factor (PDGF) may be released at sites where neutrophil proteinases may also be released, we examined the effects of neutrophil elastase and cathepsin G upon the chemotactic and mitogenic activities of PDGF. Elastase abolished the chemotactic activity of PDGF for fibroblasts but had no effect on its chemotactic activity for monocytes, or on its mitogenic activity for 3T3 cells or its capacity to bind to 3T3 cells. Cathepsin G had no effect upon the chemotactic or mitogenic activities of PDGF. In contrast, trypsin eliminated the chemotactic activity of PDGF for monocytes and fibroblasts and the mitogenic activity of PDGF. After reduction and alkylation, PDGF retained full chemotactic activity for fibroblasts and monocytes but exhibited no mitogenic activity and only limited binding to 3T3 cells. These results indicate separate domains on PDGF for fibroblast chemotactic and mitogenic activity and for monocyte and fibroblast chemotactic activity and raise the possibility that the biological activities of PDGF may be modified selectively in vivo. The findings further suggest that the majority of PDGF receptors on fibroblasts mediate mitogenic activity and that only a minority of the PDGF receptors on fibroblasts are responsible for chemotactic activity.

Hydroxysafflor yellow A attenuates high glucose-induced human umbilical vein endothelial cell dysfunction

2019 ◽  
Vol 38 (6) ◽  
pp. 685-693 ◽  
Author(s):  
S Chen ◽  
J Ma ◽  
H Zhu ◽  
S Deng ◽  
M Gu ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Growth Factor ◽  
Vascular Injury ◽  
High Glucose ◽  
Umbilical Vein ◽  
Cell Permeability ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hydroxysafflor Yellow A ◽  
Human Umbilical Vein ◽  
Nadph Oxidase 4

High glucose (HG) induces vascular injury in diabetes. Hydroxysafflor yellow A (HSYA) has been used to ameliorate ischemic cardiovascular diseases in China for many years. In the present study, we assessed whether HSYA has a potential protective role in HG-induced human umbilical vein endothelial cell (HUVEC) injury. Cell viability was determined with an 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay. Cell apoptosis was detected by fluorescein isothiocyanate/propidium iodide staining assay. The endothelial cell permeability was measured with a permeability assay. Cell adhesion molecule (CAM) expression, vascular endothelial growth factor, and basic fibroblast growth factor levels were detected with an enzyme-linked immunosorbent assay. Reactive oxygen species (ROS) formation was measured with a DCF-DA assay. Protein expression of NADPH oxidase 4 (NOX4) was measured by Western blotting. Our data indicated that HG increases HUVEC apoptosis, vascular permeability, monocyte adhesion, the level of CAMs, the formation of ROS, and NOX4 expression. Our data revealed that HG increases vascular injury, which is attenuated by HSYA. Because vascular inflammation has a key role in the development of diabetes mellitus, our results implied that HSYA is considered as a potential agent for diabetic vascular injury treatment.

Sign in / Sign up

Export Citation Format

Share Document