METHOD FOR PRODUCTION OF MATRIX-BOUND VESICLES FROM MONOLAYER CELL CULTURES AND CELL SPHEROIDS Russian patent published in 2022 - IPC C12N5/02 

FIELD: biotechnology; cell biology.

SUBSTANCE: invention relates to biotechnology and cell biology, namely to a method for the production of matrix-bound vesicles from cell cultures. For implementation of the method, first, cell cultures are decellularized, then, they are enzymatically treated to obtain a solution of a fermented extracellular matrix and matrix-bound vesicles. After that, matrix-bound vesicles are separated from fibrils of the fermented extracellular matrix by means of centrifugation. In this case, decellularization is carried out with 0.5% Triton X-100 solution in 20 mM of ammonia for at least 5 minutes per at least 1 ml of the solution per 1.5-2 million of cells or 600-800 spheroids. At the same time, enzymatic treatment is carried out with a solution of enzymes, containing dispase, collagenase I, and collagenase II, taken at concentrations of 1.5 IU/ml, 2 mg/ml, 1 mg/ml, respectively, per at least 1 ml of the solution per 1.5-2 million of cells or 600-800 spheroids.

EFFECT: invention allows for provision of high yield of matrix-bound vesicles from monolayer cell cultures and three-dimensional cell cultures in the form of spheroids, while reducing time of treatment of material.

4 cl, 1 tbl, 10 ex