Table 2.

Summary of common in vitro induction methods for ApoEVs

Method type	Specific inducer/ stimulus	Typical concentration/ duration	Origin cells (examples)	Ref.
Chemical	Staurosporine (STS)	0.5 µM for 20 h	Mesenchymal stem cells	[63]
		0.5 µM for 12 h	RAW264.7 macrophages in vitro	[64]
		0.5 µM for 3 h	Bone marrow macrophages, preosteoclasts, and mature osteoclasts in vitro	[65]
		0.5 µM for 8 h	Human ESCs, iPSCs, and umbilical cord MSCs	[66]
		0.5 µM for 12 h	Human deciduous pulp stem cells	[67]
		0.5 µM for 3 h	Neutrophils	[68]
	Alendronate (ALN)	500 µM for 24 h	Bone marrow macrophages, preosteoclasts, mature osteoclasts	[69]
	Etoposide	10 µM for 24 h	Human T lymphocytes	[70]
	Hydrogen peroxide (H₂O₂)	1 mmol/L for 12 h	Endplate chondrocyte	[71]
	Actinomycin-D	5 µg/mL for 12 h	Human T lymphocytes	[70]
	Tumor necrosis factor-α (TNF-α)	100 µM for 12 h	Human T lymphocytes	[70]
	Doxorubicin	25 µM for 48 h	Murine B16-F1 or B16-OVA cells in vitro	[27]
	Bortezomib	72 h	Human glycan-modified melanoma cell line Mel-JuSo	[72]
	Temozolomide/cisplatin	25–100 µM/20–50 µM for 24 h	Human Glioblastoma cells / LUAD cell	[73, 74]
	LPS	1 µg/mL for 24 h	RAW264.7 macrophages in vitro	[75]
	Anisomycin	5 µg/mL	HeLa cell	[76]
	Jurkat-derived microparticles (Membrane lipids)	3 × 106	RAW macrophages Human U937 cells	[77]
	CoCl2	50–100 µM for 2 d	PC12 cells	[78]
Physical	UV-B	100 mJ/cm2	Mutu I BL cells	[76]
		150 mJ/cm2	Jurkat cells	[76]
		90 mJ/cm2	Human lymphoblasts in vitro	[79]
	γ irradiation	6–12 Gy for 3–6 d	Human Glioblastoma cells in vitro	[73]
	Heat shock	56 °C for 30 min in a water bath	Jurkat cells	[70]
Physiological	Serum-free culture (Starvation)	Serum starvation for 4 h (time can vary significantly by cell type)	Monocytic leukemia cells, T cell lymphoma cells, neuronal cells, epithelial cells (human), Jurkat cells	[80, 81]