Fluidic system for producing extracellular vesicles and associated method
A fluidic system for producing extracellular vesicles from suspended producer cells, including at least one container, a liquid medium contained in the container, suspended producer cells, a liquid medium agitator, a device for controlling the speed of the agitator suitable for the growth of the suspended producer cells, wherein the device for controlling the speed of the agitator, the agitator and the shape and dimensions of the container are suitable for generating a turbulent flow of the liquid medium in the container for exerting shear stresses on the producer cells in order to carry out the production of extracellular vesicles, the Kolmogorov length of the flow being less than or equal to 50 μm.
1 . A method for ex vivo production of extracellular vesicles from suspended producer cells, comprising:
controlling the speed of an agitator adapted for the growth of the suspended producer cells,
wherein the producer cells are non-adherent cells or adherent cells detached from a culture support,
wherein a container has a shape and dimensions adapted to the generation of a turbulent flow of a liquid medium in the container to exert shear stresses on the producer cells in order to achieve the production of extracellular vesicles, the flow having a Kolmogorov length less than or equal to 50 μm in the container, the container comprising an output, the liquid medium comprising suspended producer cells, and
collecting the liquid medium comprising extracellular vesicles at the output of the container.
2 . The method according to claim 1 wherein the liquid medium is agitated for at least twenty minutes.
3 . The method according to claim 1 , wherein a separator depletes part of the liquid medium collected at the output of the container of extracellular vesicle, and wherein the part of the liquid medium is reintroduced into the container.
4 . The method according to claim 1 , wherein the method comprises a prior step of loading at least one therapeutic and/or imaging agent present in the liquid medium.
5 . The method according to claim 1 , wherein said flow allows simultaneously charging at least one therapeutic and/or imaging agent into the interior or at the membrane of the producer cells and producing the extracellular vesicles in the container.
6 . The method according to claim 1 , wherein the flow has a Kolmogorov length less than or equal to 40 μm in the container.
7 . Extracellular vesicles obtained by the method for ex vivo production of extracellular vesicles from suspended producer cells according to claim 1 .
8 . The extracellular vesicles according to claim 7 , for their use in immunotherapy, regenerative medicine, alternative or in addition to cell therapy, as a vector for delivering at least one therapeutic agent, and/or in the treatment of tumors, infectious diseases, inflammatory diseases, immunological diseases, metabolic diseases, cancer diseases, genetic diseases, degenerative diseases or diseases secondary to surgery or trauma.
9 . A method of treatment in immunotherapy, in regenerative medicine, as an alternative or in addition to cell therapy, as vectors of at least one therapeutic and/or imaging agent, and/or for the treatment of tumors, infectious diseases, inflammatory diseases, immunological diseases, metabolic diseases, cancer diseases, genetic diseases, degenerative diseases or diseases secondary to surgeries or trauma, comprising administering to a subject in need thereof the extracellular vesicles according to claim 7 .