Table 4.

Summary of the published work about the generation of mimetic EVs nanovesicles by top-down bio-nanotechnology (cell source and type of cargo are encapsulated, and main characteristics are given).

Generation technique	Precursor cell type	Type of material encapsulated	Nanovesicles characteristics	Reference
Manual extrusion over polycarbonate membrane filters	Monocytes and macrophages	Exogenous, chemotherapeutic drugs	Mean size and distribution similar to that of exosomes Exosomal protein profile similar to that of natural exosomes EE of chemotherapeutics dependent on the original amount used during extrusion High rate of drug release 100 times more that of nanovesicles than exosomes from the same number of cells Results reproducible with different cell types	[16]
Pressurization and extrusion over hydrophilic parallel microchannels in a microfluidic device	Murine embryonic stem cells	Endogenous, proteins and RNA	Average size in the exosome range Similar intracellular and membrane protein and total RNA profile to the original cells and exosomes Same ability to deliver RNA content as exosomes	[88]
Centrifugal force and extrusion over a filter with micro-size pores into a polycarbonate holder structure	Murine embryonic stem cells	Endogenous, proteins and RNA	NVs size and morphology similar to exosomes Cargo of RNA, intracellular proteins and plasma membrane proteins similar in types to exosomes Small RNA profile differs in quantity, especially in miRNA with respect to exosomes Intravesicle contain twice the concentration of natural exosomes 250 times more vesicles than naturally secreted exosomes	[87]
Slicing living cell membrane with silicon nitride blades in a microfluidic device	Murine embryonic stem cells	Exogenous, polystyrene latex beads	Generated NVs in the size range of exosomes Nanovesicle production 100 times more productive than natural exosome 30% of EE for 22 nm nanoparticles as model of exogenous material encapsulation NVs can deliver exogenous encapsulated material	[18]