scholarly journals Separation of Monocytes from Whole Human Blood

2002 ◽  
Vol 2 ◽  
pp. 1540-1543
Author(s):  
John Graham
Keyword(s):  
Human Blood ◽  
Whole Blood ◽  
Human Peripheral Blood ◽  
Preliminary Evidence ◽  
Low Density ◽  
Rapid Rate ◽  
Blood Monocytes ◽  
Peripheral Blood Monocytes ◽  
Density Barrier ◽  
Edta Anticoagulated Blood

Human peripheral blood monocytes are isolated by flotation from whole blood through a single low-density barrier prepared from OptiPrep™ at 4°C. The separation from lymphocytes depends on the more rapid rate of flotation of the monocytes because of their slightly lower density and larger size. The method works optimally only with fresh (within 2 h of drawing) EDTA-anticoagulated blood. Preliminary evidence suggests that this technique may be applicable to blood from rats.

scholarly journals Separation of Human Monocytes from a Leukocyte-Rich Plasma

2002 ◽  
Vol 2 ◽  
pp. 1646-1649 ◽  
Author(s):  
John Graham
Keyword(s):  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Rapid Rate ◽  
Human Monocytes ◽  
Blood Monocytes ◽  
Peripheral Blood Monocytes ◽  
Edta Anticoagulated Blood

Human peripheral blood monocytes are isolated by flotation from a dense leukocyte-rich plasma (LRP) through two lower-density barriers prepared from OptiPrep™. The separation from lymphocytes depends on the more rapid rate of flotation of the monocytes because of their slightly lower density and larger size. The method works optimally only with fresh (within 2 h of drawing) EDTA-anticoagulated blood.

scholarly journals Single-cell RNA sequencing reveals activated status of peripheral blood monocytes in tissue homeostasis

2021 ◽  
Author(s):  
Jiarui Lu ◽  
Junpan Luo ◽  
Junbing Guo ◽  
Jie Zeng ◽  
Xiaolei Zhang
Keyword(s):  
Single Cell ◽  
Rna Sequencing ◽  
Human Blood ◽  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Osteoclast Differentiation ◽  
Blood Monocytes ◽  
Differentiation Potential ◽  
Peripheral Blood Monocytes ◽  
Single Cell Rna Sequencing

Abstract Osteoclasts are giant multinucleated cells responsible for bone resorption, derived from myeloid cells of the monocyte/macrophage lineage. Peripheral blood monocytes have increased osteoclast differentiation potential, but only a small proportion of monocytes can differentiate into osteoclasts. The characteristics of osteoclast precursors remain unknown. The aim of this study is to explore the heterogeneity of human blood monocytes from healthy donors by use of single cell RNA sequencing (scRNA-seq), and infer the osteoclastogenic potential of the newly identified subsets with the bioinformatics approach analysis. In this study, magnetic cell separator was used to collect CD14+ monocytes from human peripheral blood, and scRNA-seq was applied as an unbiased analysis strategy to identify and analyze human blood monocyte subtypes at the single-cell resolution. Bioinformatics analysis was subsequently performed based on the differentially expressed genes (DEGs) of each cluster. Six monocyte clusters of human blood were identified, which were named of, “Initial MONO”, “Antigen presenting MONO”, “Inflamed MONO”, “Bacteriolytic MONO”, “Non-classical MONO”, and “Antivirus MONO”. Each cluster exhibited unique transcriptional profile and distinct pathway enrichments, revealing the steady-state activation of monocytes. These data provided new opportunities and insight to explore unique populations of monocytes and their different potential, and further conducive to the development of more effective anti-bone resorptive therapy.

MCP-1-stimulated monocytes preferentially utilize beta 2-integrins to migrate on laminin and fibronectin

1995 ◽  
Vol 269 (1) ◽  
pp. C60-C68 ◽  
Author(s):  
T. W. Penberthy ◽  
Y. Jiang ◽  
F. W. Luscinskas ◽  
D. T. Graves
Keyword(s):  
Extracellular Matrix ◽  
Human Peripheral Blood ◽  
Neutrophil Migration ◽  
Extracellular Matrix Proteins ◽  
Matrix Proteins ◽  
Blood Monocytes ◽  
Peripheral Blood Monocytes ◽  
Monocyte Chemoattractant Protein 1 ◽  
Monocyte Migration ◽  
Beta 2

Recruitment of monocytes to inflammatory sites involves a series of sequential attachments and detachments to extracellular matrix proteins in response to a chemoattractant gradient. In this study we compared the migration of human peripheral blood monocytes on different extracellular matrix proteins in response to monocyte chemoattractant protein-1 (MCP-1) and N-formylmethionyl-leucyl-phenylalanine. Monocytes migrated more effectively on laminin compared with other extracellular matrix proteins. In contrast, this preference was not observed with neutrophils, suggesting that the monocytes and neutrophils may have differences in their migration on extracellular matrix proteins. To study this further, function-blocking monoclonal antibodies were used to examine mechanistically whether beta 1- and beta 2-integrins were involved in monocyte migration on fibronectin or laminin in response to MCP-1. Monocyte migration on both laminin and fibronectin was blocked 100% (P < 0.05) by intact monoclonal antibody, F(ab') fragments, and F(ab')2 fragments to beta 2-integrins. We also determined that antibodies to beta 2-integrins block monocyte migration that has already been initiated. In contrast, antibody to the beta 1-integrins inhibited monocyte migration by approximately 40% (P < 0.05). Thus monocytes that express both beta 1- and beta 2-integrins require utilization of beta 2-integrins in migration on extracellular matrix proteins. The results also suggest that beta 1-integrins facilitate monocyte migration but that monocyte migration is not absolutely dependent on the interaction of beta 1-integrins with extracellular matrix proteins. In contrast, neutrophil migration is beta 2-integrin dependent and is not facilitated by beta 1-integrins.

scholarly journals Identification of normal human peripheral blood monocytes and liver as sites of synthesis of coagulation factor XIII a-chain

Blood ◽  
1987 ◽  
Vol 70 (2) ◽  
pp. 579-582
Author(s):  
LJ Weisberg ◽  
DT Shiu ◽  
PR Conkling ◽  
MA Shuman
Keyword(s):  
Peripheral Blood ◽  
Factor Xiii ◽  
Human Peripheral Blood ◽  
Cdna Probe ◽  
Coagulation Factor ◽  
Blood Monocytes ◽  
Peripheral Blood Monocytes ◽  
Normal Human ◽  
Normal Human Peripheral Blood ◽  
A Chain

Factor XIII is the fibrin-stabilizing factor that covalently cross- links fibrin monomers to form a highly organized, stable fibrin clot. The plasma form of factor XIII is a heterodimer, a2b2, consisting of two a-chains and two b-chains; the intracellular form, such as in platelets and placenta, is a dimer, a2, consisting of a-chains only. The catalytic function of factor XIII, a transglutaminase, resides in the a-chain. To address questions regarding sites of synthesis of factor XIII a-chain, an EcoRI restriction fragment from the protein- coding region of the factor XIII a-chain cDNA was used as a probe for Northern blot analysis. The cDNA probe showed hybridization with a single approximately 4.0-kilobase (kb) message in poly (A)+ mRNA prepared from normal human peripheral blood monocytes and normal human liver. The results demonstrate conclusively that factor XIII a-chains are actively synthesized in circulating monocytes and in liver. To our knowledge, these data represent the first demonstration of synthesis of any blood coagulation factor in primary uncultured and unstimulated monocytes or macrophage cells.

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