METHOD FOR DETERMINATION OF GROWTH PROPERTIES OF HUMAN BLOOD PLASMA

摘要

A method for determination of growth properties of human blood plasma includes passaging standardized cell line in a medium with a sample of examined blood with subsequent incubation and determination of growth properties. Human blood plasma drawn on an empty stomach is used as a medium, in concentration in diapason of 2.5-20%, the test cell line 4BL is introduced into plasma in 96-cup tray, 4BL cells are shifted off the surface of tray glass with Tripsin and Versene in the ratio of 1:1. After that the cells are stained with 0.2% trypan blue, calculates in Gorjaev's chamber and dissolved in cultural medium to concentration of 100 000 per ml. The cells with plasma samples were cultivated during 72 hours in CO2-incubator, after that in each cup 15 μl of solution of dye thiazoline blue tetrazolium bromide is added; cells are cultivated with the dye during 4 hours at 37 °C; the cells during testing are not washed and solution from the cups is not removed. After that 200 μl of dimethyl sulfoxide are added into the cups and the cultural medium is continuously mixed with micropipette till complete extraction of dye from the cells; growth properties of human blood plasma are determined by measuring optical density by a spectrophotometer for microtrays at 570 nm.