Organizing co mmittee

 
 

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Poster presentation 
163 
THE USE OF A NEW DENSITY GRADIENT "GLEDOL" FOR 
LYMPHOCYTE ISOLATION FROM PERIPHERAL BLOOD 
 
R.K. Rakhmanberdyeva
1
, D.M. Saidkhojaeva
1
, D.A. Rakhimov
2
, V.N. Syrov
1 
 
1) S.Yu. Yunusov Institute of the Chemistry of Plant Substances, Academy of Sciences 
Republic of Uzbekistan, 100170, Tashkent, M. Ulugbek st., 77 
2) Tashkent Institute of Chemical Technology 
 
Aqueous-insoluble polysaccharide-gledol with Mm 40 kDa was obtained from the 
seeds of 
Gleditsia triacanthos
by water-extraction. The gledol was used as part of a 
density gradient for the isolation of blood lymphocytes. For this purpose, a contrast 
agent was added to an aqueous solution of gledol to obtain a clear solution with a 
density of 1.077. 
In medical practice, a synthetic copolymer ficoll consisting of sucrose and 
epichlorohydrin is used. We carried out a comparative study of gledol with its imported 
analog, Ficoll 400 (Pharmacia, Sweden). - synthetic copolymer. 
Table 
Comparative quantitative characteristics of lymphocyte suspensions, % 
Characteristics Gradient 
characteristic cell 
suspensions, % 
Ficoll - verogranin 
Galactomannan-
verrographin 
Cell yield 
40-60 
40-60 
Erythrocyte admixture,% 
0-10 
0-10 
Other cell admixture (granulocytes: 
neutrophils, segmented cells), % 
5 
5 
Cell viability 
90-95 
90-95 
The results have shown that gledol is not inferior to ficoll in the deposition of 
immunocompetent blood cells. Gledol allows the isolation of viable peripheral blood 
lymphocytes; 35% of leucocyte suspensions can be isolated, of which 90% are 
lymphocytes, the admixture of other cells (neutrophils, segmented nuclei, etc.) is 10%. 
Gledol is used in the determination of membrane marker characteristics and functional 
activity of blood lymphocytes. 
The advantage of Gledol is that it is simple and easy to use, and that it is a domestic 
reagent of plant origin. Gledol as a diagnostic agent, is intended for immunological and 
cytological investigations in diagnostic centres and scientific research establishments.