Isolation and characterization of bovine mammary endothelial cells

Aherne, K. M.; Davis, Maria R.; Sordillo, Lorraine M.

doi:10.1007/bf00981884

Cited by 21 publications

(13 citation statements)

References 7 publications

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“…We utilized a unique culture system for the development of 2Se primary BAEC, which was based on a low-serum culture protocol previously published (1,15). Primary 2Se BAEC were subcultured in F12K medium containing 3% fetal bovine serum, antibiotics and antimycotics (100 U/ml), L-glutamine (2 mM), heparin (100 mg/ml), insulin (10 mg/ml), transferrin (5 mg/ml), and linoleic acid (1 mg/ml), and CAO ET AL.…”

Section: Preparation Of Se-deficient (2se) and Se-adequate (1se) Baecmentioning

confidence: 99%

Selenium Modulates 1-O-Alkyl-2-Acetyl-sn-Glycero-3-Phosphocholine (PAF) Biosynthesis in Bovine Aortic Endothelial Cells

Cao

Cohen

Weaver

et al. 2001

Antioxidants & Redox Signaling

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Selenium (Se) deficiency has been reported to increase platelet-activating factor (PAF) production in human endothelial cells; however, the mechanism is unclear. This study demonstrated that tumor necrosis factor-alpha (TNF-alpha) stimulated Se-deficient bovine aortic endothelial cells (BAEC) produced significantly more PAF than Se-supplemented cells. Moreover, the increase in the level of PAF was associated with enhanced activity of two anabolic enzymes in the remodeling pathway: phospholipase A2 and Lyso-PAF:acetyl-coenzyme A acetyltransferase (Lyso-PAF-AcT). In contrast, the activity of the PAF catabolic enzyme, PAF-acetylhydrolase, was not affected by Se status. Interestingly, prostacyclin, a potent vasodilator and inhibitor of platelet aggregation, inhibited the activity of Lyso-PAF-AcT and reduced the PAF production in TNF-alpha-stimulated BAEC. Therefore, we conclude that Se deficiency alters PAF production in TNF-alpha-stimulated BAEC by altering the activity of anabolic enzymes involved in the remodeling pathway partially through the inhibition of prostacyclin production.

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Section: Preparation Of Se-deficient (2se) and Se-adequate (1se) Baecmentioning

confidence: 99%

Selenium Modulates 1-O-Alkyl-2-Acetyl-sn-Glycero-3-Phosphocholine (PAF) Biosynthesis in Bovine Aortic Endothelial Cells

Cao

Cohen

Weaver

et al. 2001

Antioxidants & Redox Signaling

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“…Bovine mammary endothelial cells were harvested by trypsin digestion of the endothelium of the mammary artery, according to Aherne et al (1995) and Smits et al (1996). Endothelial cells were grown to confluence in 44% DMEM, 44% Ham's F12 nutrient mixture, 10% FBS, and 2% antibiotic-antimycotic solution at 37°C in 95% air-5% CO 2 .…”

Section: Culture Of Mammary Endothelial Cellsmentioning

confidence: 99%

Apoptosis of Bovine Neutrophils Following Diapedesis Through a Monolayer of Endothelial and Mammary Epithelial Cells

Oostveldt¹,

Paape²,

Burvenich³

2002

Journal of Dairy Science

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In a two-chamber system, isolated blood polymorphonuclear neutrophil leukocytes (PMN) were allowed to migrate (5 h, 37°C) in response to bovine complement component C5a across calfskin and rattail type I collagen-coated micropore membranes, arterial endothelial, or mammary epithelial cell monolayer on calfskin and rat-tail collagen-coated membranes, respectively. Migration through calfskin collagen-coated membranes resulted in 14.5% ± 3.4% apoptotic PMN, which was significantly higher than 6.6% ± 1.2% apoptotic nonmigrated C5a-treated PMN. The addition of an endothelial or epithelial cell monolayer to collagen-coated membranes prevented apoptosis of migrated PMN. After removing the membranes, nonmigrated (untreated and C5a treated) and migrated PMN were incubated for an additional 20 h. At this time point, 69.1% ± 4.5% and 47% ± 4.5% of PMN that have migrated through a calfskin-coated membrane and an endothelial monolayer, respectively, were apoptotic, compared with 28.2% ± 3.0% and 21.1% ± 4.5% apoptotic untreated and C5a-treated PMN, respectively; 46.9% ± 4.8% of PMN that have migrated through rat-tail-coated membranes were apoptotic compared with 14.7% ± 2.3% and 9.3% ± 1.2% apoptotic untreated and C5a-treated PMN, respectively. Migration across rat-tail collagen-coated membranes with a monolayer of epithelial cells did not affect apoptosis of migrated PMN, even after 20 h of incubation. In conclusion, migration of PMN across collagen-coated membranes (either calfskin or rat-tail collagen) induced an apoptotic response, which was

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“…Cultures were labeled with Dil-Ac-LDL as previously described [17]. Cell populations that were less than 97% BMEC (i.e., Ͼ3% contaminated with other cell types) were purified via fluorescence-activated cell sorting [17].…”

Section: Endothelial Cell Culturementioning

confidence: 99%

“…Cell populations that were less than 97% BMEC (i.e., Ͼ3% contaminated with other cell types) were purified via fluorescence-activated cell sorting [17]. Seleniumdeficient (ϪSe) and Se-supplemented (ϩSe) cells were grown according to Sordillo et al [16].…”

Section: Endothelial Cell Culturementioning

confidence: 99%

Increased neutrophil adherence and adhesion molecule mRNA expression in endothelial cells during selenium deficiency

Maddox

Aherne

Reddy

et al. 1999

Journal of Leukocyte Biology

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Leukocyte aggregation and activation on endothelial cells (EC) are important preliminary events in leukocyte migration into tissue and subsequent inflammation. Thus, an increase in leukocyte adherence has the potential to affect inflammatory disease outcome. Selenium (Se) is an integral part of the antioxidant enzyme glutathione peroxidase (GSH-Px) and plays an important role in the maintenance of the redox state of a cell. Se supplementation in the bovine has been shown to improve the outcome of acute mastitis caused by coliform bacteria, in part by enhancing the speed of neutrophil migration into the affected mammary gland. However, the mechanisms by which Se modulates neutrophil migration have not been elucidated. Therefore, an in vitro model of Se deficiency in primary bovine mammary artery EC was used to examine the impact of Se status on the adhesive properties of EC. The effect of Se on functional activities was examined by measuring neutrophil adherence to Se-deficient and Se-supplemented EC. Se-deficient EC showed significantly enhanced neutrophil adherence when stimulated with tumor necrosis factor ␣ (TNF-␣) for 4 or 24 h, interleukin-1 for 12 h, or H 2 O 2 for 20 min (P F 0.05). To determine the mechanisms underlying these changes in neutrophil adherence, the expression of EC adhesion molecules, ICAM-1, E-selectin, and P-selectin were examined at the molecular level by a competitive reverse transcription-polymerase chain reaction. Results revealed higher mRNA expression for Eselectin and ICAM-1 in Se-deficient EC stimulated with TNF-␣ for 3 and 6 h, and greater expression of P-selectin mRNA in Se-supplemented EC with 3-h TNF-␣ stimulation. These studies provide new information to establish the role of Se nutrition in the initiation of leukocyte adherence to endothelium.

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Isolation and characterization of bovine mammary endothelial cells

Cited by 21 publications

References 7 publications

Selenium Modulates 1-O-Alkyl-2-Acetyl-sn-Glycero-3-Phosphocholine (PAF) Biosynthesis in Bovine Aortic Endothelial Cells

Selenium Modulates 1-O-Alkyl-2-Acetyl-sn-Glycero-3-Phosphocholine (PAF) Biosynthesis in Bovine Aortic Endothelial Cells

Apoptosis of Bovine Neutrophils Following Diapedesis Through a Monolayer of Endothelial and Mammary Epithelial Cells

Increased neutrophil adherence and adhesion molecule mRNA expression in endothelial cells during selenium deficiency

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