US 12,467,033 B2
Production of extracellular vesicles from stem cells
Shulamit Levenberg, Moreshet (IL); Shaowei Guo, Haifa (IL); and Barak Zohar, Atlit (IL)
Assigned to TECHNION RESEARCH AND DEVELOPMENT FOUNDATION LIMITED, Haifa (IL)
Filed by TECHNION RESEARCH AND DEVELOPMENT FOUNDATION LIMITED, Haifa (IL)
Filed on Mar. 8, 2021, as Appl. No. 17/194,972.
Application 17/194,972 is a continuation in part of application No. PCT/IL2020/050641, filed on Jun. 10, 2020.
Claims priority of provisional application 62/978,317, filed on Feb. 19, 2020.
Claims priority of provisional application 62/866,702, filed on Jun. 26, 2019.
Prior Publication US 2021/0189329 A1, Jun. 24, 2021
Int. Cl. C12N 5/071 (2010.01)
CPC C12N 5/0602 (2013.01) [C12N 2513/00 (2013.01); C12N 2527/00 (2013.01)] 18 Claims
 
1. A method for producing extracellular vesicles (EVs) from an engineered tissue containing stem cells therein, the method comprising the steps of:
a) embedding a population of stem cells in at least one three-dimensional porous scaffold;
b) culturing the stem cells on and/or within the at least one three-dimensional porous scaffold for at least 6 days, to expand the scaffold and form at least one connected stem cell multi-layer engineered tissue containing stem cells and EVs therein, wherein the cultured stem cells adhere to the scaffold and/or to each other and secrete EVs;
c) providing shear stress stimulations in the range of about 5 to about 30 dyne/cm2 to said at least one engineered tissue containing the population of stem cells and EVs, wherein the population of stem cells secretes the EVs into a medium, and wherein the shear stress stimulations induce physiological changes in the cells that result in enhanced secretion of EVs compared to an engineered tissue that was not exposed to shear stress simulations;
d) collecting the medium containing the secreted EVs; and
e) isolating the secreted EVs from the medium.