Table 1.

Common cell-specific stimuli of extracellular vesicle formation and analysis.

Stimulus	Cell Type	Concentration/Incubation *	Readout
Endothelial Cells
Proinflammatory stimuli
IL1α	HUVEC	5 ng/mL 0–72 h	Annexin V binding; TF and endothelial antigen expression; procoagulant effects in vivo [35,36]
IL1β	HUVEC	10 U/mL 24 h	EV release and marker antigen expression [37]
LPS	HAEC	10 ng/mL 24 h	EV release; PS and annexin V expression [38]
PMA	HUVEC	100 ng/mL 24 h	EV release [37]
TNFα	Renal and brain microvascular and coronary artery macrovascular ECs	10 ng/mL 24 h	Marker antigen expression and annexin V binding [39]
TNFα	HUVEC	10 ng/mL 24 h	TF release by small and large EVs [40]
TNFα	HUVEC	1–100 ng/mL 6 h (blebbing) 24 h (vesiculation)	EV release and marker antigen expression; procoagulant activity in vitro [37]
TNFα	HUVEC	100 ng/mL 48 h	MP release [41]
TNFα	HBMEC	20 ng/mL 20 h	MP release [42]
TNFα	HMEC-1	100 ng/mL 48 h	MP release [43]
Procoagulant stimuli
Ca2+ ionophore A23187	HUVEC	100 µmol/L 10 min	EV release [37]
Complement C5b-9	HUVEC	2 µg in 125 µL 10 min	EV release; prothrombinase and thrombomodulin activity [44]
PAI-1	HUVEC	1–10 ng/mL 1–3 h	Procoagulant, uPAR and integrin αvβ3 expression, thrombin generation [44,45,46]
Thrombin	HUVEC	0.1–10 U/mL 24–48 h	Marker antigen expression; procoagulant MP release [37,47,48]
Thrombin	HBMEC	2 U/mL 20 h	Procoagulant MP release [42]
Proapoptotic stimuli
Serum and growth factor deprivation	Renal and brain microvascular ECs; coronary artery macrovascular ECs	N/A 24 h	Marker antigen expression and annexin V binding [39]
Staurosporine	HUVEC	100 nmol/L 2–16 h	Apoptotic MP/PS exposure; annexin V binding; TF expression; factor Xa activation [49]
Platelets
Ca2+ ionophore A23187	Human platelets	10 µM 15 min	CD62P and PS exposure, EV quantity and quality [45,46]
ABT-737	Human platelets	30 µM 90 min	CD62P and PS exposure [50]
ADP	Human platelets	60 µM 30 min	EV quantity and quality [45]
ADP	Human platelets	20 µM 1 h	EV quantity and quality [51]
Collagen	Human platelets	10 µg/mL 30 min	EV generation [45,52]
Complement C5b-9	Human platelets	2 µg in 125 µL 10 min	EV release, factor V binding [7]
CRP-XL	Human platelets	1 µg/mL 30 min	EV generation [45,52]
LPS	Human platelets	100 ng/mL 3 h	EV quantity and quality [45]
Thrombin	Human platelets	1 U/mL 15 min	CD62P and PS exposure [50]
Thrombin	Human platelets	1 U/mL 30 min	EV generation [45,52]
Thrombin and CaCl2	Human platelets	0.25 U/mL 5 min	EV quantity and quality [53]
TRAP6	Human platelets	60 µM 30 min	EV quantity and quality [45]
Monocytes
Ca2+ ionophore A23187	THP-1	10 mM 5 min	TF+ MP release [37,54]
LPS	THP-1	5 µg/mL 4 h	MP release [55]
LPS	THP-1	1 µg/mL 1 h (RT)	MP release [56]
LPS	THP-1	1 µg/mL 5 h (RT)	TF+ MP release [54]

* Incubation at 37 °C, if not stated otherwise.