METHODICAL ASPECTS OF USING RAT RBL-2H3 BASOPHILS FOR DEGRANULATION STUDY

DOI: https://doi.org/None

M.A. Goryacheva, K.L. Kryshen, PhD, A.E. Katelnikova, E.G. Akulova, M.N. Makarova, DMSci, V.G. Makarov, DMSci, professor Scientific and Production Association «HOUSE OF PHARMACY», Building 245, u.s.Kuzmolovskiy, Saint-Petersburg, 188663, Russian Federation

Introduction. Mediator release from mast cells or basophils is the key event in many acute allergic and inflammatory processes. Rat basophilic leukemia cell line RBL-2H3 cells is widely used as a model for studying mast cells degranulation. Aim of the study. The aim of the study was the working off the conditions of the induction of rat basophilic leukemia cell line RBL-2H3 degranulation (selection of the optimal inductor, the time of exposition and the concentration range). Material and methods. Cells were stimulated by the known inductors: compound 48/80 and calcium ionophore A23187 in a wide concentration range. The incubation time of cell culture in the presence of the inductors was 10, 25 and 45 minutes. Cell degranulation was assessed by beta-hexosaminidase enzyme activity in supernatants and cells lysates. Results. The differences in the level of degranulation depending on the concentration and incubation time with the inductors were observed. The dependence on the concentration of both substances was typically bell-shaped. In total, the induction of degranulation by calcium ionophore А23187 was more effective and resulted in a higher percentage of beta-hexosaminidase enzyme net release compared with compound 48/80. The mechanisms of the compound 48/80 and the calcium ionophore A23187 action as an inductor of mast cell degranulation were discussed.
Keywords: 
mast cells, RBL-2H3, degranulation, compound 48/80, calcium ionophore A23187

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