METHOD FOR PRODUCING GROWTH ADDITIVE BASED ON THROMBOCYTE LYSATE FROM PLATELET OF DONORS TO MEDIUM FOR BUILD-UP OF CELLULAR WEIGHT OF STEM, PROGENITOR, DIFFERENTIATED AND TUMOR CELLS

摘要

FIELD: biotechnology.;SUBSTANCE: invention relates to a method for producing a growth additive to a medium for cultivating human cells from platelet of donors. The present method includes rating of a platelet sample to a predetermined platelet concentration of 1.75×10⁹ c/ml by centrifugation of the platelet at 3130 g for 40 minutes at 20°C and resuspension of the precipitate in the supernatant of a predetermined volume to a specified platelet concentration. Three-time temperature lysis of the normalized sample by freezing for a day to down -80°C and defrosting at +37°C. Deposition of cell debris by centrifugation at 3130 g for 40 minutes at 20°C, collection of supernatant obtaining an individual sample from thrombocyte lysate with its subsequent microscopic control while increasing ×200 for the availability in the field of vision of no more than 3 thrombocyte fragments. While standardization is carried out by pooling equal in volume individual samples of lysate, derived from platelet of not less than 12 donors of both bid teams, and the obtained thrombocyte lysate is centrifuged at 3130 g for 40 minutes at 20°C, with the subsequent ultrafiltration in filters of 0.45 µm or 0.22 mcm. The obtained growth additive is divided into aliquots of the required volume and frozen down to -80°C.;EFFECT: invention allows for obtaining standardized samples of nonxenogenic growth additive for the cultivation of different types of human cells with the functional activity of the samples, determined by the speed of growth of the test cultures.;3 dwg, 8 tbl, 2 ex