Culture media-based selection of endothelial cells, pericytes, and perivascular-resident macrophage-like melanocytes from the young mouse vestibular system

Abstract. Inflammatory and oxidative stress in endothelial cells are implicated in the pathogenesis of premature atherosclerosis in diabetes. To determine whether high-dextrose concentrations induce the expression of pro-inflammatory cytokines, human coronary artery endothelial cells (HCAEC) were exposed to either 5.5 or 27.5 mM dextrose for 24-hours and interleukin-1β (IL-1β), interleukin-2 (IL-2), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor α (TNF α) levels were measured by enzyme immunoassays. To determine the effect of antioxidants on inflammatory cytokine secretion, cells were also treated with α-tocopherol, ascorbic acid, and the glutathione peroxidase mimetic ebselen. Only the concentration of IL-1β in culture media from cells exposed to 27.5 mM dextrose increased relative to cells maintained in 5.5 mM dextrose. Treatment with α-tocopherol (10, 100, and 1,000 μM) and ascorbic acid (15, 150, and 1,500 μM) at the same time that the dextrose was added reduced IL-1β, IL-6, and IL-8 levels in culture media from cells maintained at 5.5 mM dextrose but had no effect on IL-1β, IL-6, and IL-8 levels in cells exposed to 27.5 mM dextrose. However, ebselen treatment reduced IL-1β, IL-6, and IL-8 levels in cells maintained in either 5.5 or 27.5 mM dextrose. IL-2 and TNF α concentrations in culture media were below the limit of detection under all experimental conditions studied suggesting that these cells may not synthesize detectable quantities of these cytokines. These results suggest that dextrose at certain concentrations may increase IL-1β levels and that antioxidants have differential effects on suppressing the secretion of pro-inflammatory cytokines in HCAEC.

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Natriuretic Peptides Regulate the Expression of Tissue Factor and PAI-1 in Endothelial Cells

Thrombosis and Haemostasis ◽

10.1055/s-0037-1614861 ◽

1999 ◽

Vol 82 (11) ◽

pp. 1497-1503 ◽

Cited By ~ 20

Author(s):

Hajime Tsuji ◽

Hiromi Nishimura ◽

Haruchika Masuda ◽

Yasushi Kunieda ◽

Hidehiko Kawano ◽

...

Keyword(s):

Endothelial Cells ◽

Tissue Factor ◽

Plasminogen Activator ◽

Natriuretic Peptide ◽

Culture Media ◽

Tissue Type ◽

Dependent Manner ◽

Ang Ii ◽

Plasminogen Activator Inhibitor 1 ◽

Pai 1

SummaryIn the present study, we demonstrate that brain natriuretic peptide (BNP) and C-type natriuretic peptide (CNP) interact with angiotensin II (Ang II) in regulative blood coagulation and fibrinolysis by suppressing the expressions of both tissue factor (TF) and plasminogen activator inhibitor-1 (PAI-1) induced by Ang II. The expressions of TF and PAI-1 mRNA were analyzed by northern blotting methods, and the activities of TF on the surface of rat aortic endothelial cells (RAECs) and PAI-1 in the culture media were respectively measured by chromogenic assay.Both BNP and CNP suppressed the expressions of TF and PAI-1 mRNA induced by Ang II in a time- and concentration-dependent manner via cGMP cascade, which suppressions were accompanied by respective decrease in activities of TF and PAI-1. However, neither the expression of tissue factor pathway inhibitor (TFPI) nor tissue-type plasminogen activator (TPA) mRNA was affected by the treatment of BNP and CNP.

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Selection of filamentous fungi that are resistant to the herbicides atrazine, glyphosate and pendimethalin

Acta Scientiarum Agronomy ◽

10.4025/actasciagron.v43i1.51656 ◽

2021 ◽

Vol 43 ◽

pp. e51656

Author(s):

Nara Priscila Barbosa Bravim ◽

Anatércia Ferreira Alves ◽

José Fábio França Orlanda ◽

Patricia Barbosa Rodrigues Silva

Keyword(s):

Filamentous Fungi ◽

Mycelial Growth ◽

Culture Medium ◽

Agricultural Soils ◽

Culture Media ◽

Fusarium Verticillioides ◽

Fungal Growth ◽

Relative Growth ◽

Maximum Inhibition ◽

Selection Of

The objective of the present study was to isolate fungi from agricultural soils and evaluate fungal growth in culture medium contaminated with atrazine, glyphosate and pendimethalin. Filamentous fungi were isolated from agricultural soils and cultured in a modified culture medium containing 0, 10, 20, 50, and 100 μg mL-1 atrazine, glyphosate and pendimethalin for 14 days at 28°C. The fungi that presented optimal and satisfactory growth were plated in Sabouraud culture medium with 4% dextrose and containing the herbicides at concentrations of 0, 10, 20, 50, and 100 μg mL-1 for seven days at 28°C. The mean mycelial growth values were submitted to analysis of variance and the Tukey test (p < 0.05%) for comparison and relative growth determination, and maximum inhibition rates were calculated. The isolated fungi Aspergillus fumigatus, Fusarium verticillioides and Penicillium citrinum were shown to be resistant to atrazine, glyphosate and pendimethalin. F. verticillioides showed higher mean mycelial growth in the culture media contaminated with atrazine and glyphosate than the other two fungi. In the culture medium contaminated with pendimethalin, F. verticillioides, and A. fumigatus presented the highest mean mycelial growth values.

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Selection of the optimal culture medium for cultivation Fusarium sporotrichioides Sherb

PROCEEDINGS OF V INTERNATIONAL SCIENTIFIC CONFERENCE “CURRENT STATE, PROBLEMS AND PROSPECTS OF THE DEVELOPMENT OF AGRARIAN SCIENCE” ◽

10.33952/2542-0720-2020--5-9-10-3 ◽

2020 ◽

Author(s):

I.L. Astapchuk ◽

◽

N.A. Marchenko ◽

G.V. Yakuba ◽

A.I. Nasonov ◽

...

Keyword(s):

Growth Rate ◽

Growth And Development ◽

Maximum Degree ◽

Culture Medium ◽

Culture Media ◽

High Variability ◽

Fusarium Sporotrichioides ◽

Cultural Characteristics ◽

Colony Diameter ◽

Selection Of

The influence of various culture media on the growth, morphological and cultural characteristics of the fungus F. sporotrichioides was studied. Ten culture media were used in our research. A comparative study of the growth rate of the F. sporotrichioides mycelium made it possible to identify two media that are the most suitable for the cultivation and identification of this species, namely carrot and tomato agar. We took into account such criteria as ensuring the maximum degree of sporulation, rapid growth and development of mycelium (the 7th day), colony diameter (71–78 mm), as well as the ease of preparation. Nirenberg culture medium can be used to obtain a large number of conidia of the fungus. Because of the high variability of cultural characteristics of F. sporotrichioides, we recommend using different composition of media.

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TFPIβ is the GPI-anchored TFPI isoform on human endothelial cells and placental microsomes

Blood ◽

10.1182/blood-2011-10-388512 ◽

2012 ◽

Vol 119 (5) ◽

pp. 1256-1262 ◽

Cited By ~ 37

Author(s):

Thomas J. Girard ◽

Elodee Tuley ◽

George J. Broze

Keyword(s):

Endothelial Cells ◽

Tissue Factor ◽

Feedback Inhibition ◽

Culture Media ◽

Factor Viia ◽

Factor Xa ◽

Gpi Anchor ◽

C Terminus ◽

Before And After ◽

Tissue Factor Pathway

Abstract Tissue factor pathway inhibitor (TFPI) produces factor Xa-dependent feedback inhibition of factor VIIa/tissue factor-induced coagulation. Messages for 2 isoforms of TFPI have been identified. TFPIα mRNA encodes a protein with an acidic N-terminus, 3 Kunitz-type protease inhibitor domains and a basic C-terminus that has been purified from plasma and culture media. TFPIβ mRNA encodes a form in which the Kunitz-3 and C-terminal domains of TFPIα are replaced with an alternative C-terminus that directs the attachment of a glycosylphosphatidylinositol (GPI) anchor, but whether TFPIβ protein is actually expressed is not clear. Moreover, previous studies have suggested that the predominant form of TFPI released from cells by phosphatidylinositol-specific phospholipase C (PIPLC) treatment is TFPIα, implying it is bound at cell surfaces to a separate GPI-anchored coreceptor. Our studies show that the form of TFPI released by PIPLC treatment of cultured endothelial cells and placental microsomes is actually TFPIβ based on (1) migration on SDS-PAGE before and after deglycosylation, (2) the lack of a Kunitz-3 domain, and (3) it contains a GPI anchor. Immunoassays demonstrate that, although endothelial cells secrete TFPIα, greater than 95% of the TFPI released by PIPLC treatment from the surface of endothelial cells and from placental microsomes is TFPIβ.

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Metabolomic profiling of culture media by near infrared spectroscopy as an adjunct to morphology for selection of a single day 3 embryo to transfer in ivf: a double blind randomised trial

Fertility and Sterility ◽

10.1016/j.fertnstert.2011.07.018 ◽

2011 ◽

Vol 96 (3) ◽

pp. S3 ◽

Cited By ~ 2

Author(s):

C.G. Vergouw ◽

D.C. Kieslinger ◽

E.H. Kostelijk ◽

P.G. Hompes ◽

R. Schats ◽

...

Keyword(s):

Infrared Spectroscopy ◽

Near Infrared Spectroscopy ◽

Near Infrared ◽

Culture Media ◽

Randomised Trial ◽

Double Blind ◽

Metabolomic Profiling ◽

Selection Of ◽

Day 3 Embryo

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Abstract P324: Unhealthy Diet and Air Pollution Compromise Human Cardiovascular Cell Functions: A First Mechanistic Analysis

Circulation Research ◽

10.1161/res.109.suppl_1.ap324 ◽

2011 ◽

Vol 109 (suppl_1) ◽

Author(s):

Niloofar Ale-Agha ◽

Nicole Buechner ◽

Ulrich Sydlik ◽

Klaus Unfried ◽

Joachim Altschmied ◽

...

Keyword(s):

Endothelial Cells ◽

Cardiovascular Diseases ◽

Carbon Black ◽

Ultrafine Particles ◽

Culture Media ◽

Density Lipoprotein ◽

Cell Functions ◽

Unhealthy Diet ◽

Cardiovascular Aging ◽

The Impact

Diet and pollution are environmental factors known to compromise ″healthy cardiovascular aging″. The molecular consequences of the permanent burden for aging of the cardiovascular system are unknown, since they have never been examined in primary, adult human cells. Therefore, this study investigates the impact of unhealthy diet on aging-related signaling pathways of human, primary cardiovascular cells and of airborne particles on human endothelial cells, as several studies demonstrated that ultrafine particles can enter the circulation and thus may interact with endothelial cells directly. Nutrition health reports have shown that the diet in industrialized countries contains more than 100 mg/dl low density lipoprotein (LDL) and a too high fraction of monosaccharides, especially fructose, which is metabolized insulin-independently. Both components have been shown to increase the risk for cardiovascular diseases. To simulate unhealthy diet we supplemented cell culture media of human, primary endothelial cells (EC), smooth muscle cells (SMC) and cardiomyocytes (CM) with 100 mg/dl LDL and replaced 1/3 of the glucose with fructose for one week. This treatment did not induce cell death in any of the cell types. However, we observed increased senescence, loss of endothelial nitric oxide synthase and increased nuclear localization of Foxo3 in EC, increased proliferation in SMC and hypertrophy in CM. With respect to pollution we have used ultrafine carbon black particles (ufCB), one of the major constituents of industrial and exhaust emissions, in concentrations our vessels are constantly exposed to. These concentrations of ufCB are non-toxic and non-inflammatory for EC. Despite these missing immediate effects, ufCB dramatically reduced the S-NO content, a marker for NO-bioavailability in EC and increased reactive oxgen species formation. As a consequence, ufCB dramatically increased senescence of EC after two weeks. Thus, unhealthy diet and a high burden of ultrafine carbon black nanoparticles, to which we are exposed every day, seem to induce a ″cardiovascular aging″ phenotype and can lead to severe cardiovascular diseases.

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Venular endothelial cells from bovine heart

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.1988.254.6.h1211 ◽

1988 ◽

Vol 254 (6) ◽

pp. H1211-H1217 ◽

Cited By ~ 9

Author(s):

M. E. Schelling ◽

C. J. Meininger ◽

J. R. Hawker ◽

H. J. Granger

Keyword(s):

Endothelial Cells ◽

In Vitro Model ◽

Protein Transport ◽

Culture Conditions ◽

Support Cell ◽

Vitro Model ◽

Microvascular Cells ◽

Selection Of ◽

Coronary Angiogenesis

Coronary venular endothelial cells were isolated by a bead-perfusion technique that allowed the selection of endothelial cells from venules of a specific size. Culture conditions for the microvascular cells were established. Cells grew well in supplemented Dulbecco's modified Eagle's medium. The effect of various substrata on the proliferation of the venular endothelial cells was determined. Matrigel, gelatin, and fibronectin supported high levels of proliferation. Cell shape was correlated with ability of the substratum to support cell proliferation. Cells exhibiting a broad, flattened morphology achieved high levels of proliferation. The formation of vessel meshworks by the coronary venular endothelial cells provides an in vitro model for the study of coronary angiogenesis. Confluent monolayers of these cells can be utilized to examine mechanisms of water and protein transport across coronary venules.

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Studies of Multimerin in Human Endothelial Cells

Blood ◽

10.1182/blood.v91.4.1304 ◽

1998 ◽

Vol 91 (4) ◽

pp. 1304-1317 ◽

Cited By ~ 18

Author(s):

Catherine P. M. Hayward ◽

Elisabeth M. Cramer ◽

Zhili Song ◽

Shilun Zheng ◽

Roxanna Fung ◽

...

Keyword(s):

Endothelial Cells ◽

Von Willebrand Factor ◽

Culture Media ◽

Cellular Membrane ◽

Human Endothelial Cells ◽

Early Passage ◽

Endothelial Cell Cultures ◽

Dense Core Granules ◽

Von Willebrand ◽

Willebrand Factor

Abstract Multimerin is a novel, massive, soluble protein that resembles von Willebrand factor in its repeating, homomultimeric structure. Both proteins are expressed by megakaryocytes and endothelial cells and are stored in the region of platelet α-granules resembling Weibel-Palade bodies. These findings led us to study the distribution of multimerin within human endothelial cells. Multimerin was identified in vascular endothelium in situ. In cultured endothelial cells, multimerin was identified within round to rod-shaped, dense-core granules, some of which contained intragranular, longitudinally arranged tubules and resembled Weibel-Palade bodies. However, multimerin was found primarily in different structures than the Weibel-Palade body proteins von Willebrand factor and P-selectin. After stimulation with secretagogues, multimerin was observed to redistribute from intracellular structures to the external cellular membrane, without detectable accompanied secretion of multimerin into the culture media. In early passage endothelial cell cultures, multimerin was associated with extensive, fibrillary, extracellular matrix structures, in a different distribution than fibronectin. Although multimerin and von Willebrand factor are stored together in platelets, they are mainly found within different structures in endothelial cells, indicating that there are tissue-specific differences in the sorting of these soluble, multimeric proteins.

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