Human pluripotent stem cell-derived extracellular vesicles: From now to the future

Table 1 Human pluripotent stem cell-derived extracellular vesicles: Methods of isolation and vesicle size

Ref.	Culture medium	EV collection time	EV isolation method	EV mean size (nm)
Bobis-Wozowicz et al[37], 2015	Essential 8™ medium	NI (cells in 70%-90% confluency)	DC + UC	146
Ju et al[50], 2017	PSCeasy medium (Cellapy)	24 h	DC + UC	122, 132
Zhou et al[51], 2017	mTeSR™-1 medium	NI (cells in 60%-90% confluency)	DC + 0.22 μm filter + UC	101
Ding et al[52], 2018	mTeSR™-1 medium	48 h	DC + UC	103.1
Kaur et al[53], 2018	Essential 8™ Flex medium	48 h	DC + UC or miR-CURY™ Exosome Isolation Kit (Exiqon A/S)	100-200
Kobayashi et al[54], 2018	DMEM-F12 + NEAA, 200 mM L-gln, KSR, 0.1 M BME	2-3 d before passage	MagCapture Exosome Isolation Kit PS (Wako)	100
Oh et al[55], 2018	Essential 8 medium	Daily, from day 2 to day 5	0.45 μm filter + ExoQuick-TC kit	85.8
Peng et al[56], 2018	mTeSR™-1 medium	24 h (cells in about 80% confluency)	MV: DC + 16500 g, 1 h; EXO: DC + 120000 g, 2 h	MV = 200-600; EXO = 40-80
Saito et al[57], 2018	mTeSR™-1 medium	NI	DC + concentration in 100-KDa filter + MagCapture™ Exosome Isolation Kit PS	179
Chen et al[58], 2019	mTeSR™-1 medium	NI	DC + UC	50-150
Liu et al[59], 2019	Essential 8™ medium	Daily, for 3-5 d	DC + concentration in 100-kDa filter + SEC	150
Marzano et al[60], 2019	mTeSR™-1 medium	Daily, for 4 d	0.22 μm filter + concentration in 100-kDa filter + Total Exosome Isolation Reagent (Thermo Fisher) or DC + UC	about 240
Povero et al[61], 2019	NI	24-48 h	DC + UC	300-400
Sun et al[62], 2019	mTeSR™-1 medium	48 h	DC + concentration in 100-kDa filter + Exosome Isolation Kit (PureExo) + UC + 0.22 μm filter	70-100 (cell-dependent)
Zhu et al[63], 2019	mTeSR™-1 medium	48h (cells 80%-90% confluency)	DC + 0.22 μm filter + UC	70.2
Collino et al[64], 2020	mTeSR™-1 medium	24 h	DC + UC	119
Hu et al[65], 2021	ncEpic hPSC medium	NI	DC + 0.22 μm filter + UC	72.4 ± 21.3
Kurtzwald-Josefson et al[66], 2020	DMEM/F12 Ham 1:1 + 20% KSR, 1% NEAA, 1% L-gln, 0.2% BME, 4 ng/mL rhFGF basic	24 h (cells in about 80% confluency)	Total exosome isolation reagent (Thermo Fisher Scientific)	115 ± 7
Liu et al[67], 2020	mTeSR™-1 medium	NI	DC + UC	50-75
Wang et al[68], 2020	PGM1 medium	NI	DC + 0.22 μm filter + UC	30-120
Andrade et al[69], 2021	mTeSR™-1 mediuma	Daily, for 4-5 days	TFF with or without subsequent UC	103-109
Ashok et al[39], 2021	StemMACS medium with 10 μM ROCK inhibitor and 0.2% Pluronic F68	Days 3, 4, and 5 prior to differentiation	DC + 0.22 μm filter + UC + SG	50
Hu et al[65], 2021	ncEpic hPSC medium	NI	DC + 0.22 μm filter + UC	-100
Karnas et al[70], 2021	Essential 8™ medium	NI	DC + UC	215.7
Ke et al[71], 2021	Exo-depleted FBS	48 h	MV: DC + 16500 g, 60 min; Exo: DC + 120000 g, 120 min	MV = 200-600; Exo = 40-80
Luo et al[49], 2021	DMEM/F12 + KSR (0.5%, 2.5%, 5%, or 20%)	Daily, for 5 d	DC + 0.45 μm filter + concentration in 10-kDa filter + 0.22 μm filter + UC or ExoQuick-TC kit (SystemBioscience)	187.8, 168.2
Saito et al[46], 2021	StemFit AK-03N medium (Ajinomoto)	NI	15000 × g, 30 min + 0.22 μm filter + UC	70
Wang et al[72], 2021	mTeSR™-1 medium	NI	DC + UC	120-140
Xia et al[73], 2021	Nuwacell hiPSC/hESCs medium	24 h	DC + 0.22 μm filter + UC	50-150
Bi et al[74], 2022	ncTarget medium (Nuwacell. Ltd, China)	24 h (cells in about 80% confluency)	DC + UC + 0.22 μm filter + UC	hESC = 133.1; hIPSC = 157.7
Gu et al[75], 2022	mTeSR™-1 medium	NI	0.45 μm filter + concentration in 100-kDa filter + GC + 0.22 μm filter + UC	143.5
Gupta et al[76], 2022	StemFlex™ medium	48 h	DC + one-step sucrose cushion UC	123.6 ± 60
Hsueh et al[77], 2023	StemFlex™ medium	48 h	DC + UC	136.8
Li et al[78], 2023	ncEpic hPSC medium	NI	DC + UC	74.70 ± 20.77
Li et al[79], 2022	mTeSR™-1 medium	NI	DC + 0.22 μm filter + UC	50.75-105.7
Pan et al[80], 2022	mTeSR™-1 medium	24 h (cells in about 80% confluency)	DC + UC	142.2 ± 64.1

DC: Differential centrifugation; EXO: Exosome; FBS: Fetal bovine serum; MV: Microvesicle; NI: Not informed; SEC: Size exclusion chromatography; SG: Sucrose gradient; TFF: Tangential flow filtration; UC: Ultracentrifugation; hPSC: Human pluripotent stem cell; hESC: Human embryonic stem cells; hIPSC: Human induced pluripotent stem cell; KSR: KnockOut™ Serum Replacement.