METHOD OF MESENCHYMAL STEM CELL ISOLATION FROM BONE MARROW BEFORE CULTIVATION Russian patent published in 2011 - IPC C12N5/775 

FIELD: medicine.

SUBSTANCE: method involves preparing a bone marrow (BM) aspirate after a iliac bone puncture, analysing a BM sample for the presence of infectious agents and bacterial contaminations. Then the BM sample is placed in heparin water 15 ml produced by dilution of heparin 5 ml in physiologic saline 200 ml. A growth medium is prepared by adding fetal bovine serum 5 ml to the concentration not less than 10% preheated to temperature 37°C to the MEM-cc or Advanced Stem Cell Media medium 45 ml. After dilution of the BM sample in Hanks's solution in 1.5-2 times, the BM suspension is layered on ficoll in the ratio 30 ml of the suspension to 15 ml of ficoll, and the solution is centrifuged with acceleration 800G for 30 min; 2 upper layers are collected with lower of these layers shaped as a white ring with pure leukocyte isolation. These two layers are transferred to a test tube containing the MEM-wasps or Advanced Stem Cell Media culture medium 7 ml, and the produced solution is centrifuged with acceleration 400G for 5 min; a growth medium 5-10 ml is added to a precipitate, and the prepared suspension is pipetted on 1-2 bottles that is followed with stem cell cultivation.

EFFECT: invention allows higher uniformity and quality of producing the higher-purity MSC population.