	US 12,239,671 B2
	Extracellular vesicles derived from human liver stem cells (HLSCs)
Paul Robbins, Minneapolis, MN (US); Giovanni Camussi, Turin (IT); and Maria Beatriz Herrera Sanchez, Turin (IT)
Assigned to UNICYTE EV AG, Oberdorf (CH)
Appl. No. 17/617,036
Filed by UNICYTE EV AG, Oberdorf (CH)
PCT Filed Jun. 9, 2020, PCT No. PCT/EP2020/065986 § 371(c)(1), (2) Date Dec. 7, 2021, PCT Pub. No. WO2020/249567, PCT Pub. Date Dec. 17, 2020.
Claims priority of application No. 19179220 (EP), filed on Jun. 10, 2019.
Prior Publication US 2022/0226389 A1, Jul. 21, 2022 Prior Publication US 2023/0037275 A2, Feb. 2, 2023
Int. Cl. A61K 35/407 (2015.01); A61P 39/00 (2006.01)

CPC A61K 35/407 (2013.01) [A61P 39/00 (2018.01)]

3 Claims

1. A method of manufacturing a pharmaceutical preparation of extracellular vesicles (EVs) derived from human liver stem cells, comprising the steps of:

isolating EVs from multiple preparations of a body fluid or from the conditioned medium of a cell culture;

preparing one or more samples from the isolated EVs at a predetermined concentration of EVs;

testing the activity of each EVs sample in a potency test measuring the reduction of cellular senescence wherein senescence is measured in a SA-β-galactosidase-based cellular senescence assay using test senescent cells not treated with EVs as a control;

selecting the preparations in which the reduction of cellular senescence measured exceeds a predetermined threshold; and

pooling two or more selected preparations, wherein

preparations selected at a concentration of 2.5×10⁷EVs/ml reduce the senescence of a population of test senescent cells by at least 10%.