METHOD FOR PRODUCING CYTOKINES FROM UMBILICAL BLOOD THROMBOCYTES AS SUBSTRATE FOR DEVELOPING DRUGS FOR HUMANS AND ANIMALS Russian patent published in 2019 - IPC A61K35/19 

FIELD: medicine.

SUBSTANCE: invention refers to medicine, and can be used for producing cytokines from umbilical blood thrombocytes. Method involves three stages: at the first stage, umbilical blood is sampled from different donors, Stabisol solution is administered in each packet with donor blood in amount of 20 % of the blood volume of the packet, each blood pack is centrifuged in a smooth program change mode, as a result of which blood is separated into thrombocyte enriched plasma and erythrocytes. Said program consists of the following modes: acceleration to 540 RCF for 5 minutes; acceleration to 900 RCF for 2 minutes; braking to 300 RCF and rotation for 2 minutes; braking up to 100 RCF and rotation for 1.3 minutes with subsequent disconnection of the centrifuge, then removing the erythrocytes from the packs and recycling them. At the second stage, all donor OTP packets are combined and all thrombocytes are concentrated, for this purpose the produced thrombocyte enriched plasma is transferred into the appropriate number of 50 ml-type falcon test tubes, centrifugation is carried out for concentration of thrombocytes in mode of 15 minutes at rate of 4000g, as a result of centrifugation a platelet concentrate and lean plasma are obtained. Oblong plasma is taken for use as a solvent, leaving 5 ml of plasma at the bottom of the test tubes, obtained thrombocyte concentrates of different donors are combined in one test tube with volume of 50 ml and diluted with depleted plasma to volume 50 ml or the solvent used is normal saline, if further plasma proteins need to be excluded; obtained thrombocyte concentrate pool is re-centrifuged in 60-minute mode at rate of 50g for removal of erythrocyte and leukocyte residues. After centrifugation, the supernatant fluid in amount of 45–48 ml is transferred into new 50 ml test tube, and 1 ml of the solution is sent to a laboratory for platelet count on an automated haematological analyzer. At the third stage, the obtained thrombocyte concentrate with a known amount of thrombocytes per ml or mcl is subjected to a storage/activation procedure, wherein the falcon with thrombocyte concentrate is lowered into a cryogenic storage of a dry type for 15 minutes at a temperature of -196 degrees Celsius, frozen product is stored in a freezing chamber at a temperature of -80 degrees Celsius, where it is stored for up to 3 years. Further activation of the product is carried out before immediate use by rapid thawing, for this purpose the falcon is placed in a water bath with a temperature of +37 degrees Celsius until complete thawing of the product, and if necessary to obtain a product with concentration of cytokines/thrombocytes less than in a frozen product, the end product is dissolved in the corresponding number of times, wherein the initial concentration of thrombocytes is used.

EFFECT: disclosed method allows to prepare a cocktail of cytokines for its further use as a substrate for preparing medicinal preparations, the method is economical and enables to produce a product with cytokine concentration of up to 1 million/mcl.

1 cl