METHOD FOR PRODUCING THREE-DIMENSIONAL CELL STRUCTURES IN MAMMALS USING 2,5-DIHYDROXYBENZOIC ACID POLYMERS Russian patent published in 2021 - IPC C12N15/00 C12N5/02 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology and specifically to production of spheroids by treating cells with polymers of 2,5-DHBC from attached cell cultures of mammals. Method involves culturing cells in a standard growth medium to a sub-monolayer stage with subsequent detachment from the substrate, resuspending and transfer of cells into growth medium containing polymers of 2,5-DHBC (hereinafter molecular weight of polymers of 40 kDa) in concentration from 50 mcg/ml to 400 mcg/ml, and cells incubation till formation of spheroids. Cells can be cultured in a standard growth medium until cells reach high density with subsequent replacement of growth medium on growth medium with addition of polymers of 2,5-DGBK in concentration from 50 mcg/ml to 400 mcg/ml and separation of cells from substrate by means of mechanical action and incubating the obtained cell aggregates without separation into separate cells in a growth medium with the addition of polymers of 2,5-DHBC in concentration of 50 mcg/ml to 400 mcg/ml until formation of spheroids. Single cell can be cultured on multilayer plates with low-adhesive coating to form clones with subsequent replacement of growth medium with medium, containing polymers of 2,5-DHBC in concentration from 50 mcg/ml to 400 mcg/ml, and separation of cells from substrate by mechanical methods and incubation of clones in suspension in presence of polymers in concentration from 50 mcg/ml to 400 mcg/ml to formation of spheroids, consisting of cells that are descendants of a single cell and therefore representing a clone of genetically identical cells.

EFFECT: invention enables to obtain three-dimensional structures (spheroids) both from a group of cells, and by cloning a single cell.

1 cl, 9 ex, 4 dwg