Influence of different anticoagulants on monocyte procoagulant functions and monocyte-platelet aggregates formation.

Author(s): Basavaraj MG, Østerud B, Hansen JB

Publication: J Thromb Haemost, 2012, Vol. 10, Page 1698-702

PubMed ID: 22712695 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of anticoagulant type on platelet counts, PBMC yields, monocyte procoagulant functions, and MPA formation in blood from healthy volunteers. Whole blood and freshly isolated PBMC were used to determine the percentages of MPA and MAPA, and cryopreserved PBMC that were thawed in a 37 degrees C water bath were used for other analyses.

   Summary of Findings:

   Specimens collected in sodium heparin tubes had lower platelet counts than specimens in sodium citrate or EDTA tubes (p<0.05 and p<0.001, respectively). PBMC yields and the proportions of CD14-positive monocytes were highest in specimens collected in EDTA, followed by specimens collected in citrate, and lastly those collected in heparin. The type of anticoagulant used had no effect on the baseline or LPS stimulated expression of TF mRNA, TF or TFPI antigen expression, or TF activity in PBMC. Under resting conditions, PBMC from heparinized whole blood had less TFPIalpha mRNA than those from citrated whole blood (p<0.05). Upon LPS stimulation for 6 h, PBMC from citrated whole blood had more TFPIalpha mRNA than those form EDTA whole blood (p<0.05). While whole blood had a lower proportion of MPA and MAPA than isolated PBMC, there was a higher ratio between MAPA and MPA in whole blood than PBMC. In both whole blood and PBMC, there was higher MPA (CD14-positive/CD42a-positive) expression than MAPA (CD14-positive/CD62P-positive). Among whole blood specimens, collection with EDTA led to lower MPA and MAPA than collection with citrate or heparin.

   Biospecimens

   • Bodily Fluid - Blood
   • Cell - Blood

   Preservative Types

   • Frozen
   • None (Fresh)

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   Cell count/volume	Flow cytometry
   RNA	Real-time qRT-PCR
   Protein	Western blot
   Protein	Enzyme assay
   Protein	Fluorescent microscopy

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Aliquots and Components	Blood and blood products	Peripheral blood mononuclear cells Whole blood
   Analyte Extraction and Purification	Incubation duration/condition	LPS stimulation for 0 h 2 h 4 h 5 h 6 h 8 h 12 h
   Biospecimen Preservation	Type of fixation/preservation	Frozen None (fresh)
   Biospecimen Acquisition	Anticoagulant	Sodium citrate Sodium heparin Potassium EDTA

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