Table 1. Overview of the different... | American Society of Hematology

Table 1.

Overview of the different isolation and marker strategies, used in MM studies

Vesicle type	Isolation method	Protocol	Size†	Markers	Reference
exo	Isolation kits, ∗∗∗also used UC	Exoquick	≤150 nm	CD63, CD81,+Alix∗ or +Tsg101∗∗	22,26,46,49,45∗,51,54∗∗,63∗∗∗
				CD63, CD81, Tsg101, syntenin	34
				Hsp70, flotilin 1	60
				Hsp70, CD63, CD9,+Alix∗ or +LAMP2∗∗ calreticulin negative	56,67∗,61∗∗
				HspP70/90, Alix, Tsg101, flotilin 1, calreticulin negative	23
				Hsp70/90, flotilin 1, CD63,+Alix∗ calreticulin negative	42,52,65∗
				CD63, CD9, flotilin 1, calreticulin negative	77
		ExoEasy kit	≤150 nm	Tsg101	59
			?	CD63, CD81	57,58
		Total exosome isolation kit	≤150 nm	CD63, Hsp70	50
	Differential centrifugation	1	≤100 nm	LAMP1, Alix, galectin negative	78
		2	≤100 nm	Alix, CD63, calnexin negative no explicit markers∗	31,32,35∗,36∗
		3	100 nm	Hsp70, CD63, Tsg101	25
		4	≤150 nm	Rab5, LAMP2, CD63, GM130 negative	80
		5	≤150 nm	Hsp70, Tsg101, CD81, CD63, calreticulin negative, no markers∗ only Hsp70∗∗	44∗,47∗∗,48
	∗Also used exoquick for patient material			CD63, flotilin 1, clathrin only CD63∗	62,72,73∗
		6	150 nm	Tsg101, Alix, GM130, and α-tubulin negative	33
	∗Also used different kits for patient material	7	≤150 nm	Tsg101, CD81	68∗
		8	≤200 nm	CD63, no markers∗	27,66∗,69
		9	50-350 nm	Alix, CD9, LAMP1	40
	UC	100 000g, 1 h	≤100 nm	CD63, Hsp70	37
		120 000g, 2 h	150 nm	CD63, Alix, Tsg101, Rab5	53
MV	Centrifugation	13 000g	≤200 nm	No markers	24
MV		16 000g	?	No markers	28
		10	99% 100 nm, 1%> 300 nm	No markers	29
		11	100 nm to 1μm	Alix, Tsg101, CD63, CD9	39
		12	≤200 nm	No markers	68
ApoEV		13	150 nm	CD63, CD9, flotilin 1, Alix, syntenin, annexin	79
MV or exo?	Concentration via centrifugal filters		150 nm	Alix, CD81	76

Vesicle type	Isolation method	Protocol	Size†	Markers	Reference
exo	Isolation kits, ∗∗∗also used UC	Exoquick	≤150 nm	CD63, CD81,+Alix∗ or +Tsg101∗∗	22,26,46,49,45∗,51,54∗∗,63∗∗∗
				CD63, CD81, Tsg101, syntenin	34
				Hsp70, flotilin 1	60
				Hsp70, CD63, CD9,+Alix∗ or +LAMP2∗∗ calreticulin negative	56,67∗,61∗∗
				HspP70/90, Alix, Tsg101, flotilin 1, calreticulin negative	23
				Hsp70/90, flotilin 1, CD63,+Alix∗ calreticulin negative	42,52,65∗
				CD63, CD9, flotilin 1, calreticulin negative	77
		ExoEasy kit	≤150 nm	Tsg101	59
			?	CD63, CD81	57,58
		Total exosome isolation kit	≤150 nm	CD63, Hsp70	50
	Differential centrifugation	1	≤100 nm	LAMP1, Alix, galectin negative	78
		2	≤100 nm	Alix, CD63, calnexin negative no explicit markers∗	31,32,35∗,36∗
		3	100 nm	Hsp70, CD63, Tsg101	25
		4	≤150 nm	Rab5, LAMP2, CD63, GM130 negative	80
		5	≤150 nm	Hsp70, Tsg101, CD81, CD63, calreticulin negative, no markers∗ only Hsp70∗∗	44∗,47∗∗,48
	∗Also used exoquick for patient material			CD63, flotilin 1, clathrin only CD63∗	62,72,73∗
		6	150 nm	Tsg101, Alix, GM130, and α-tubulin negative	33
	∗Also used different kits for patient material	7	≤150 nm	Tsg101, CD81	68∗
		8	≤200 nm	CD63, no markers∗	27,66∗,69
		9	50-350 nm	Alix, CD9, LAMP1	40
	UC	100 000g, 1 h	≤100 nm	CD63, Hsp70	37
		120 000g, 2 h	150 nm	CD63, Alix, Tsg101, Rab5	53
MV	Centrifugation	13 000g	≤200 nm	No markers	24
MV		16 000g	?	No markers	28
		10	99% 100 nm, 1%> 300 nm	No markers	29
		11	100 nm to 1μm	Alix, Tsg101, CD63, CD9	39
		12	≤200 nm	No markers	68
ApoEV		13	150 nm	CD63, CD9, flotilin 1, Alix, syntenin, annexin	79
MV or exo?	Concentration via centrifugal filters		150 nm	Alix, CD81	76

exo, exosomes; MV, microvesicles; UC, ultracentrifugation.

∗,∗∗,∗∗∗Asterisks refer to which marker or isolation method is exceptionally used (in addition to the generally listed markers) in the corresponding reference.

†

Size was measured via transmission electron microscope, and/or nanoparticle tracking analysis, dynamic light scattering, or flow cytometry. Different differential centrifugation protocols were used: (1) 300g (5 minutes), 1200g (20 minutes), 10 000g (30 minutes), 100 000g (60 minutes); (2) 300g (5 minutes), 3000g (15 minutes), 10 000g (30 minutes), 100 000g (90 minutes); (3) 800g (30 min), 16 000g (45 min), 100 000g (2 hours); (4) 300g (10 minutes), 4000g (20 minutes), 100 000g (4 hours + 1 hours); (5) 300g (10 minutes), 2000g (20 minutes), 100 000g (30/70 minutes); (6) 450g (5 minutes), 16 000g (15 minutes), 120 000g (20 minutes); (7) 500g (5 minutes), 2000g (10 minutes), 10 000g (30 minutes), 100 000g (60 minutes); (8) 2000g (30 minutes), 20 000g (70 minutes), 170 000g (90 minutes); (9) 300g (10 minutes), 12 000g (45 minutes), 100 000g (90 minutes). Different centrifugation protocols were used: (10) 1500g (15 minutes), 20 000g (60 minutes); (11) 200g, (5 minutes), 1500g (30 minutes), 16 000g (60 minutes); (12) 500g (5 minutes), 2000g (10 minutes), 10 000g (30 minutes); (13) 800g (10 minutes), 2000g (10 minutes), 16 000g (30 minutes).

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