Table 3. Studies evaluating the... | American Society of Hematology

Table 3.

Studies evaluating the procoagulant activity of NETs ex vivo

Reference	Species	Method of neutrophil isolation	Method/duration of NET induction	Coagulation assay	Main conclusions
Oemcke et al 2009⁵³	Human	Gradient centrifugation	Glucose oxidase (60 min) or IL8 (60 min) or M1 protein-fibrinogen complexes (60 min)	Chromogenic assay for PK hydrolysis in plasma	NETs bind FXII and HK, and promote PK hydrolysis in plasma
Fuchs et al 2010⁹⁸	Human	Gradient centrifugation	PMA (4 h), glucose oxydase (1U/ml, 4 h)	Electron and fluorescent microscopy after flowing plasma, suspension of platelets or whole blood over NETs seeded in a flow chamber or on glass cover slide	NETs bind platelets and promote platelet activation and aggregation.
					NETs bind RBCs, VWF, fibrinogen, and fibronectin.
					NETs promote resistance to tPA-mediated fibrinolysis
Gould et al 2014¹⁰¹	Human	Gradient centrifugation	PMA 150 min	Thrombin generation in PPP or PRP	NETs enhance thrombin generation
Noubouossie et al 2017²⁴	Human	Magnetic bead-based negative selection	PMA, calcium ionophore and LPS.	Contact system in buffer	NETS do not trigger contact system activation in buffer, do not trigger thrombin generation in plasma, and do not amplify low TF-initiated TG in PFP.
Noubouossie et al 2017²⁴	Human	Magnetic bead-based negative selection	4 h	Thrombin generation in PFP and PRP
Ivanov et al 2017⁵⁷	Human	Gradient centrifugation	PMA: 90 min	Thrombin generation in plasma	NETs shorten lag time of thrombin generation.
Wang et al 2018¹⁰²	Mouse	Gradient centrifugation	PMA: 3 h	Thrombin generation in plasma	NETs form complexes with microvesicles to enhance TG in plasma

Reference	Species	Method of neutrophil isolation	Method/duration of NET induction	Coagulation assay	Main conclusions
Oemcke et al 2009⁵³	Human	Gradient centrifugation	Glucose oxidase (60 min) or IL8 (60 min) or M1 protein-fibrinogen complexes (60 min)	Chromogenic assay for PK hydrolysis in plasma	NETs bind FXII and HK, and promote PK hydrolysis in plasma
Fuchs et al 2010⁹⁸	Human	Gradient centrifugation	PMA (4 h), glucose oxydase (1U/ml, 4 h)	Electron and fluorescent microscopy after flowing plasma, suspension of platelets or whole blood over NETs seeded in a flow chamber or on glass cover slide	NETs bind platelets and promote platelet activation and aggregation.
					NETs bind RBCs, VWF, fibrinogen, and fibronectin.
					NETs promote resistance to tPA-mediated fibrinolysis
Gould et al 2014¹⁰¹	Human	Gradient centrifugation	PMA 150 min	Thrombin generation in PPP or PRP	NETs enhance thrombin generation
Noubouossie et al 2017²⁴	Human	Magnetic bead-based negative selection	PMA, calcium ionophore and LPS.	Contact system in buffer	NETS do not trigger contact system activation in buffer, do not trigger thrombin generation in plasma, and do not amplify low TF-initiated TG in PFP.
Noubouossie et al 2017²⁴	Human	Magnetic bead-based negative selection	4 h	Thrombin generation in PFP and PRP
Ivanov et al 2017⁵⁷	Human	Gradient centrifugation	PMA: 90 min	Thrombin generation in plasma	NETs shorten lag time of thrombin generation.
Wang et al 2018¹⁰²	Mouse	Gradient centrifugation	PMA: 3 h	Thrombin generation in plasma	NETs form complexes with microvesicles to enhance TG in plasma

LPS, lipopolysaccharide; PMA, phorbol myristate acetate; tPA, tissue-type plasminogen activator.