METHOD OF CULTIVATION OF AMNIOTIC LIQUID CELLS Russian patent published in 2017 - IPC C12N5/00 

FIELD: biotechnology.

SUBSTANCE: invention is a method of cultivation of amniotic liquid cells comprising adding amniotic liquid of the nutrient medium to the cells and incubating in vials, several changes of the nutrient medium followed by treatment of cells with colchicine, detaching the cells from the bottom of the vial, hypotonic treatment and treatment with a fixative, where "Amniomax" or "Amniocar" nutrient medium are used as a nutrient medium in the ratio of the initial hallmark: medium, equal to 1.4:1, the first change of the medium is carried out 5 days from the beginning of cultivation with the pretreatment of cells with Hank's solution at a ratio of 2.3:1, the subsequent changes of the medium are performed every 2 days, then in 6-8 days from the beginning of cultivation trypsinization is performed with 0.05% trypsin-EDTA solution for 2-3 minutes at 37°C, the first trypsinization is carried out in one of the vials of the cell culture with the most active growth, the next day trypsinization is carried out in the second vial with a medium rise, a day later - in the third vial with minimal growth, treatment of cells is carried out with colchicine with a concentration of 1 µg/ml for 2.5 hours at 37°C in the presence of 5% CO₂, the detachment of the cells from the bottom of the vial is carried out with 0.05% of trypsin-EDTA solution for 2-3 minutes at 37°C, hypotonic treatment of the cells is carried out with an aqueous solution of embryonic calf serum (2:1) for 20 minutes at 37°C, and fixation of cellsis carried out with a solution containing ethanol and glacial acetic acid at a ratio of 3:1.

EFFECT: acceleration of the method and increasing the yield of cultured cells.

1 dwg, 1 tbl, 2 ex