Flow cytometric analysis of tissue factor (TF) expression on stimulated monocytes — comparison to procoagulant activity of mononuclear blood cells

1990 ◽  
Vol 61 (6) ◽  
pp. 375-378 ◽  
Author(s):  
Th. Luther ◽  
C. Flössel ◽  
V. Hietschhold ◽  
R. Koslowski ◽  
M. Müller
Keyword(s):  
Tissue Factor ◽  
Blood Cells ◽  
Flow Cytometric Analysis ◽  
Procoagulant Activity ◽  
Flow Cytometric ◽  
Mononuclear Blood Cells

scholarly journals Flow Cytometric Analysis of Phenotypes of Canine Red Blood Cells with FITC-Labeled Clerodendron Trichotomum Lectin.

1994 ◽  
Vol 56 (3) ◽  
pp. 593-595 ◽  
Author(s):  
Reiko USUI ◽  
Junko HIROTA ◽  
Toshinori OMI ◽  
Sadahiko IWAMOTO ◽  
Shigenori IKEMOTO
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Flow Cytometric Analysis ◽  
Flow Cytometric ◽  
Clerodendron Trichotomum

scholarly journals Flow cytometric analysis of Notch1 and Jagged1 expression in normal blood cells and leukemia cells

2012 ◽  
Vol 4 (3) ◽  
pp. 397-400 ◽  
Author(s):  
ERIKO KANAMORI ◽  
MAI ITOH ◽  
NAOKO TOJO ◽  
TAKATOSHI KOYAMA ◽  
NOBUO NARA ◽  
...  
Keyword(s):  
Blood Cells ◽  
Flow Cytometric Analysis ◽  
Leukemia Cells ◽  
Normal Blood ◽  
Flow Cytometric

P-37 Flow cytometric analysis of cytosolic and mitochondrial ferritins in immature red blood cells from patients with myelodysplastic syndrome

2005 ◽  
Vol 29 ◽  
pp. S38
Author(s):  
M.G. Della Porta ◽  
L. Malcovati ◽  
M. Maffioli ◽  
E. Travaglino ◽  
S. Levi ◽  
...  
Keyword(s):  
Myelodysplastic Syndrome ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Flow Cytometric Analysis ◽  
Flow Cytometric

Microfluidic structures for flow cytometric analysis of hydrodynamically focussed blood cells fabricated by ultraprecision micromachining

Lab on a Chip ◽  
2009 ◽  
Vol 9 (7) ◽  
pp. 972 ◽  
Author(s):  
A. Kummrow ◽  
J. Theisen ◽  
M. Frankowski ◽  
A. Tuchscheerer ◽  
H. Yildirim ◽  
...  
Keyword(s):  
Blood Cells ◽  
Flow Cytometric Analysis ◽  
Flow Cytometric

scholarly journals Purification Platelets from Mouse Blood with Sickle Cell Disease Using Iohexol Gradient Medium

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 4888-4888
Author(s):  
Md Nasimuzzaman
Keyword(s):  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Blood Cells ◽  
Flow Cytometric Analysis ◽  
Cell Disease ◽  
Mouse Blood ◽  
Simple Method ◽  
Flow Cytometric ◽  
Microscopic Evaluation ◽  
Gradient Medium

Platelets are purified from whole blood to study their functional properties, which should be free from red blood cells (RBC), white blood cells (WBC), and plasma proteins. Since RBC and WBC contain significantly more RNA and proteins than platelets, the presence of even a small number of these cells can interfere with transcriptomic and proteomic analyses of RNA and proteins derived from platelets. Several protocols have described the isolation of platelets from human, dog, rat, and non-human primate by various methods. Some of the methods require multiple steps such as collection of platelet-rich plasma by centrifugation, filtration by separation column, negative selection of platelets with RBC- and WBC-specific antibody conjugated to magnetic beads, and so on, which are time-consuming and may degrade platelets and their contents. Moreover, mouse blood with sickle cell disease contains a significant level of fragments of RBC which should be removed from the platelet preparation. However, the mouse yields a relatively smaller volume of blood, which makes it difficult to purify platelets. If the same small volume of gradient medium and blood samples are used, the platelet layer cannot be clearly separated from RBC-WBC layer after centrifugation. We describe a simple method for purification of platelets from mouse blood with sickle cell disease using three-fold more iohexol gradient medium relative to blood sample volume and centrifugation in a swinging bucket rotor at 400 x g for 20 min at 20 °C. The platelet layer is collected and centrifuged again at 200 x g for 20 min at 20 °C to remove the residual fragments of RBC. The recovery/yield of the purified platelets were 10-17%, and the purified platelets were in a resting state, which did not contain any significant number of RBC and WBC. The purified platelets were activated with thrombin indicating their viability. We confirmed that the purified platelets were sufficiently pure using flow cytometric and microscopic evaluation. Although flow cytometric analysis of purified platelet from sickle cell disease mice showed a few RBC events after staining with anti-TER119 antibody, the microscopic study did not show any intact RBC or larger fragments indicating that these are smaller fragments of RBC which do not interfere with the biochemical and functional studies. This method can be used for purification of platelets from the blood of other species and disease models as well. Disclosures No relevant conflicts of interest to declare.

Increased Levels of Tissue Factor mRNA in Mononuclear Blood Cells of Patients with Primary Antiphospholipid Syndrome

1999 ◽  
Vol 82 (12) ◽  
pp. 1578-1582 ◽  
Author(s):  
Chari López-Pedrera ◽  
Francisco Velasco ◽  
María Aguirre ◽  
Antonio Torres ◽  
María Cuadrado ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Antiphospholipid Syndrome ◽  
Blood Cells ◽  
Polymerase Chain Reaction Amplification ◽  
Primary Antiphospholipid Syndrome ◽  
Mrna Accumulation ◽  
Cardiolipin Antibodies ◽  
History Of ◽  
Mononuclear Blood Cells ◽  
Normal Controls

SummaryAntiphospholipid antibodies (aPL) may stimulate tissue factor (TF) expression in cultured endothelial cells and monocytes, but there are discrepancies as to the expression of TF in the patients with antiphospholipid syndrome (APS). By using reverse transcription and polymerase chain reaction amplification, we have analysed TF mRNA accumulation in freshly isolated mononuclear blood cells (MBC) of 14 patients with primary APS (PAPS) and six normal controls. TF mRNA accumulation was low or absent in uncultured MBC from all normal controls, but was elevated in uncultured MBC from nine of the patients as well as in normal MBC incubated with 100 ng/ml lipopolysaccharide (LPS). Mean levels of TF mRNA, as measured by densitometry, were higher in MBC from patients (N = 14) than in those from controls (N = 6, P = 0.009), and in MBC from patients with a history of thrombosis (N = 9) than in those from patients without thrombosis (N = 5, P = 0.02). Uncultured MBC of patients with thrombosis accumulated TF mRNA at similar levels to LPS-treated normal MBC. Increased levels of TF mRNA were found in eight of ten patients with conventional aPL (ie, anti-cardiolipin antibodies [aCL] and/or lupus anticoagulant [LA]) and littleif any accumulation of TF mRNA was observed in three of four patients without aPL at the time of study. These data strongly suggest that circulating monocytes of many patients with PAPS are subjected to an up-regulated TF expression that may well explain their prothrombotic state. Although the presence or absence of TF mRNA in MBC was associated with, respectively, the presence or absence of conventional aPL in 11 of the 14 patients studied, our study cannot exclude the involvement of factors other than aCL or LA in inducing TF expression.

Interleukin-6 Directly Modulates Stem Cell Factor-Dependent Development of Human Mast Cells Derived From CD34+Cord Blood Cells

Blood ◽  
1999 ◽  
Vol 94 (2) ◽  
pp. 496-508 ◽  
Author(s):  
Tatsuya Kinoshita ◽  
Nobukuni Sawai ◽  
Eiko Hidaka ◽  
Tetsuji Yamashita ◽  
Kenichi Koike
Keyword(s):  
Mast Cell ◽  
Stem Cell ◽  
Mast Cells ◽  
Cord Blood ◽  
Interleukin 6 ◽  
Blood Cells ◽  
Stem Cell Factor ◽  
Flow Cytometric Analysis ◽  
Flow Cytometric ◽  
Cord Blood Cells

In the present study, we attempted to clarify the effects of interleukin-6 (IL-6) on the growth and properties of human mast cells using cultured mast cells selectively generated by stem cell factor (SCF) from CD34+ cord blood cells. The addition of IL-6 to cultures containing mast cells resulted in a substantial reduction of the number of progenies grown by SCF in the liquid culture. This IL-6–mediated inhibition of mast cell growth may be due in part to the suppression at the precursor level, according to the results of a clonal cell culture assay. Moreover, a flow cytometric analysis showed that the cultured mast cells grown in the presence of SCF+IL-6 had decreased c-kit expression. The exposure of cultured mast cells to SCF+IL-6 also caused substantial increases in the cell size, frequency of chymase-positive cells, and intracellular histamine level compared with the values obtained with SCF alone. The flow cytometric analysis showed low but significant levels of expression of IL-6 receptor (IL-6R) and gp130 on the cultured mast cells grown with SCF. The addition of either anti–IL-6R antibody or anti-gp130 antibody abrogated the biological functions of IL-6. Although IL-4 exerted an effect similar to that of IL-6 on the cultured mast cells under stimulation with SCF, the results of comparative experiments suggest that the two cytokines use different regulatory mechanisms. Taken together, the present findings suggest that IL-6 modulates SCF-dependent human mast cell development directly via an IL-6R-gp130 system.

scholarly journals Automated Flow Cytometric Analysis of Blood Cells in Cerebrospinal Fluid

2004 ◽  
Vol 121 (5) ◽  
pp. 690-700 ◽  
Author(s):  
Marthe W. Aune ◽  
Joanne L. Becker ◽  
Carlo Brugnara ◽  
William Canfield ◽  
David M. Dorfman ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Blood Cells ◽  
Flow Cytometric Analysis ◽  
Flow Cytometric

scholarly journals Comparative gene expression profiling of in vitro differentiated megakaryocytes and erythroblasts identifies novel activatory and inhibitory platelet membrane proteins

Blood ◽  
2006 ◽  
Vol 109 (8) ◽  
pp. 3260-3269 ◽  
Author(s):  
Iain C. Macaulay ◽  
Marloes R. Tijssen ◽  
Daphne C. Thijssen-Timmer ◽  
Arief Gusnanto ◽  
Michael Steward ◽  
...  
Keyword(s):  
Expression Profiling ◽  
Blood Cells ◽  
Transmembrane Domain ◽  
Flow Cytometric Analysis ◽  
Structural Features ◽  
Low Doses ◽  
Flow Cytometric ◽  
Bioinformatical Analysis ◽  
G Protein Coupled

AbstractTo identify previously unknown platelet receptors we compared the transcriptomes of in vitro differentiated megakaryocytes (MKs) and erythroblasts (EBs). RNA was obtained from purified, biologically paired MK and EB cultures and compared using cDNA microarrays. Bioinformatical analysis of MK–up-regulated genes identified 151 transcripts encoding transmembrane domain-containing proteins. Although many of these were known platelet genes, a number of previously unidentified or poorly characterized transcripts were also detected. Many of these transcripts, including G6b, G6f, LRRC32, LAT2, and the G protein–coupled receptor SUCNR1, encode proteins with structural features or functions that suggest they may be involved in the modulation of platelet function. Immunoblotting on platelets confirmed the presence of the encoded proteins, and flow cytometric analysis confirmed the expression of G6b, G6f, and LRRC32 on the surface of platelets. Through comparative analysis of expression in platelets and other blood cells we demonstrated that G6b, G6f, and LRRC32 are restricted to the platelet lineage, whereas LAT2 and SUCNR1 were also detected in other blood cells. The identification of the succinate receptor SUCNR1 in platelets is of particular interest, because physiologically relevant concentrations of succinate were shown to potentiate the effect of low doses of a variety of platelet agonists.

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