Demonstration of Antithrombin III in Fresh and Cultured Endothelial Cells from Human Umbilical Cord

1979 ◽  
Author(s):  
Vivian Chan ◽  
T.K. Chan
Keyword(s):  
Endothelial Cells ◽  
Umbilical Cord ◽  
Antithrombin Iii ◽  
Cultured Cells ◽  
Active Role ◽  
Human Umbilical Cord ◽  
Thrombosis Research ◽  
Natural Inhibitor ◽  
Immunofluorescent Technique ◽  
Radio Immunoassay

We have shown by immunofluorescent technique that the distribution of antithrombin III (ATIII) in human tissues was concentrated around the microvasculature of the lungs and kidneys, as well as veins and small arteries of other organs (liver and spleen). It would seem that ATIII is stored and/or synthesized in the endothelial cells similar to Factor VIII-RAG and Plasminogen Activator. Endothelial cells were isolated from human umbilical cord by collagenase and cultured according to Chemethod described by Shearn etal (1977). In freshly isolated endothelial cells, ATIII could be demonstrated by indirect immunof1uorescent technique and radio immunoassay confirmed the presence of 14.8 ng per 106 cells. After 7 days’ culture, the supernatant from 106 cells contained about 15 ng and the cultured cells (106) contained 16.9 ng ATIII. The presence of ATIII in cultured cells was also confirmed by the positive immunofluorescence. Hence the endothelial cells play an active role in preventing thrombosis by the synthesis and liberation of ATIII, the major natural inhibitor of the intrinsic pathway of Coagulation.Reference: Shearn S.A., Peake I.R., Ciddings J.C., Humphrys J. and Bloom A.L. Thrombosis Research, 11, 43, 1977.

scholarly journals Venous and Arterial Endothelial Cells from Human Umbilical Cords: Potential Cell Sources for Cardiovascular Research

2021 ◽  
Vol 22 (2) ◽  
pp. 978
Author(s):  
Skadi Lau ◽  
Manfred Gossen ◽  
Andreas Lendlein ◽  
Friedrich Jung
Keyword(s):  
Endothelial Cells ◽  
Umbilical Cord ◽  
Membrane Integrity ◽  
Cell Types ◽  
Cell Number ◽  
Human Umbilical Cord ◽  
Cardiovascular Research ◽  
Cardiovascular Devices ◽  
Arterial Endothelial Cells

Although cardiovascular devices are mostly implanted in arteries or to replace arteries, in vitro studies on implant endothelialization are commonly performed with human umbilical cord-derived venous endothelial cells (HUVEC). In light of considerable differences, both morphologically and functionally, between arterial and venous endothelial cells, we here compare HUVEC and human umbilical cord-derived arterial endothelial cells (HUAEC) regarding their equivalence as an endothelial cell in vitro model for cardiovascular research. No differences were found in either for the tested parameters. The metabolic activity and lactate dehydrogenase, an indicator for the membrane integrity, slightly decreased over seven days of cultivation upon normalization to the cell number. The amount of secreted nitrite and nitrate, as well as prostacyclin per cell, also decreased slightly over time. Thromboxane B2 was secreted in constant amounts per cell at all time points. The Von Willebrand factor remained mainly intracellularly up to seven days of cultivation. In contrast, collagen and laminin were secreted into the extracellular space with increasing cell density. Based on these results one might argue that both cell types are equally suited for cardiovascular research. However, future studies should investigate further cell functionalities, and whether arterial endothelial cells from implantation-relevant areas, such as coronary arteries in the heart, are superior to umbilical cord-derived endothelial cells.

scholarly journals Dexamethasone and lidocaine suppress eosinophilopoiesis from umbilical cord blood cells

2020 ◽  
Vol 18 (1) ◽  
Author(s):  
Masato Muraki ◽  
Hirohito Kita ◽  
Gerald J. Gleich
Keyword(s):  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Cultured Cells ◽  
Eosinophil Infiltration ◽  
Human Umbilical Cord ◽  
Dependent Manner ◽  
Eosinophil Granule Proteins ◽  
Granule Proteins ◽  
Eosinophil Granule

Abstract Background Eosinophils play an important role in allergic inflammation. Glucocorticosteroids have been used as an anti-inflammatory medication for inflammatory diseases involving eosinophil infiltration. Some effect of nebulized lidocaine has been reported when treating certain patients with asthma, which is also an inflammatory disease. The goal of this study was to examine the effects of dexamethasone and lidocaine on eosinophil proliferation and differentiation using a model of human umbilical cord blood mononuclear cells (UCMC) cultured with IL-5. Methods UCMC were cultured with IL-5 (5 ng/mL) for 4 weeks. The effects of dexamethasone and lidocaine on the number and morphology of eosinophilic cells were visualized with Wright-Giemsa and cyanide-resistant peroxidase stains. Moreover, the effect on eosinophil-derived neurotoxin (EDN) and eosinophil peroxidase (EPX) contents in cultured cells were evaluated using radioimmunoassay. Results The number of eosinophilic cells and EDN and EPX content in cultured cells increased in a time-dependent manner in the presence of IL-5. Dexamethasone treatment slightly decreased the number of eosinophilic cells in one week, but this effect was lost in 2–4 weeks. Macrophages in cultured UCMC treated with dexamethasone contained more eosinophil granule proteins. Both EDN and EPX content in cultured cells were reduced by dexamethasone. Lidocaine decreased the number of eosinophilic cells and reduced both EDN and EPX contents in cultured cells. Conclusions Dexamethasone suppressed the production of eosinophil granule proteins and may also induce apoptosis of eosinophils, while lidocaine suppresses eosinophilopoiesis.

INTERACTION BETWEEN CULTURED ENDOTHELIAL CELLS AND ABNORMAL ANTITHROMBIN III "TOYAMA"

1987 ◽  
Author(s):  
N Sakuragawa ◽  
S Saitoh ◽  
K Takahashi
Keyword(s):  
Endothelial Cells ◽  
Room Temperature ◽  
Antithrombin Iii ◽  
Cultured Cells ◽  
Binding Activity ◽  
Endothelial Cell Culture ◽  
Antithrombin Activity ◽  
Thrombin Activity ◽  
Cultured Endothelial Cells ◽  
At Iii

Purpose: Abnormal antithrombin III(AT-III)Toyama showed non-affinity to heparin and heparinoid to show loss of immediate antithrombin activity. On the endothelial cells, there are heparinoids including heparan sulfate. We investigated on the interaction between cultured endothelial cells and abnormal AT-III"Toyama" from the viewpoint of antithrombin activity.Materials and methods: (1) Endothelial cell culture:^125I-labelled normal and abnormal AT-III were placed on the washed endothelial cultured cells in 0.2 ml of RPMI-1640 medium for 15 min at 37°C. The medium was suctioned off and the cell layer was washed with Hank's balanced salt solution. The cells were incubated with 1 ml of heparin(3 ug/ml) for 15 min at 4°C. The radioactivity in the supernatant was counted, and represented AT-III which bound to the cells surface. (2) Antithrombin activity: 0.23 ml of thrombin solution^ U/ml) and 0.03 ml of normal or abnormal AT-III plasma were mixed, and incubated on the cultured cell surface for 5 min at room temperature. The residual thrombin activity was assayed by 0.3 ml of the substrate (S-2238) solution(0.8mM)for 5 min. After these procedures,2 ml of 2% citric acid solution was added to stop the reaction, and 0D(405 nm) was recorded.Results: Abnormal AT-III showed reduced binding-activity to cultured cells to one fifth compared with normal AT-III, and the residual thrombin activity in the abnormal was higher compared with that in normal plasma.Conclusion: Abnormal AT-III showed less binding activity to the cultured endothelial cells, and less thrombin neutralizing activity to show thrombogenic tendency.

scholarly journals Yellow fever virus strains Asibi and 17D-204 infect human umbilical cord endothelial cells and induce novel changes in gene expression

Virology ◽  
2005 ◽  
Vol 342 (2) ◽  
pp. 167-176 ◽  
Author(s):  
Svetlana F. Khaiboullina ◽  
Albert A. Rizvanov ◽  
Michael R. Holbrook ◽  
Stephen St. Jeor
Keyword(s):  
Gene Expression ◽  
Endothelial Cells ◽  
Umbilical Cord ◽  
Yellow Fever ◽  
Yellow Fever Virus ◽  
Fever Virus ◽  
Human Umbilical Cord ◽  
Virus Strains

Induction of Human Umbilical Cord Blood Mesenchymal Stem Cells into Nerve-Like Cells by Salvia Miltiorrhiza.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4227-4227
Author(s):  
Lian Ma ◽  
Xueyong Feng ◽  
Liye Yang ◽  
Minjie Luo ◽  
Qingdong Xie ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Cultured Cells ◽  
Salvia Miltiorrhiza ◽  
Neural Induction ◽  
Human Umbilical Cord Blood ◽  
Human Umbilical Cord

Abstract Salvia miltiorrhiza is a traditional Chinese herb with thousands of years history. Composite salvia injection (CSI), a widely used administration in China nowadays, consists of Salvia miltiorrhiza, notoginseng, radix and borneo. It can promote blood flow, but also regulate qi to alleviate pain. So it is frequently used for angina, coronary heart disease and chest discomfort. Recently, some laboratory studies show that CSI may have the effect of cell induction.. In this study, we aim to investigate its neural induction. Objective: To investigate the differentiation-inducing effects of salvia miltiorrhiza on umbilical cord blood mesenchymal stem cells. Methods: The immunophenotypes of human umbilical cord blood-derived mesenchymal stem cells(MSCs) were determined by flow-cytometry. Salvia miltiorrhiza was adapted to induce the differentiation of the primary cultured cells of umbilical cord blood and MSCs. The results were compared with nerve growth factor(NGF) and ganglioside(GM1). The differentiated and undifferentiated cells were identified with immunocytochemistry. Results: In primary culture, certain cells proliferated and formed cell clusters. These cells were either fibroblast-like or round in shape with various sizes. The positive rates for MSCs phenotypes CD29, CD44, CD59 of primary culture cells were 10.7%,37.27% and 66.67%, respectively. Salvia miltiorrhiza could induce the primary cultured cells of umbilical cord blood to express neural phenotypes. After passage, the MSCs could be maintained in vitro for extended periods with stable population doubling. The positive rates for CD29, CD44, CD59 of human umbilical cord blood-derived MSCs were 40.2%,70.5% and 95.4%, respectively. Salvia miltiorrhiza could induce the MSCs to express three kinds of proteins: nestin(the marker of neuronal precursor stem cells), β-TubulinIII and neurofilament (NF) (the marker of neuronal phenotypes) and glial fibrillary acidic protein(GFAP, the marker of astrocyte). Besides, NGF and GM1 could also induce the same MSCs to nerve-like cell, which could express the same markers: β-TubulinIII, NF and GFAP. In comparison NGF and GM1, the induction effect of salvia miltiorrhiza demonstrated the higher rate of expressing neuron phenotypes at a faster speed. Conclusion: The human MSCs could be isolated from human umbilical cord blood. Salvia miltiorrhiza can induce the MSCs to differentiate into nerve-like cells, therefore, might act as a potential drug for neural induction.

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