Эффективность различных методов выращивания вируса африканской чумы свиней в гемопоэтических клетках

Abstract

For African swine fever virus (ASFV) reproduction little publications are available, in particular, those concerning cultivation technologies. Moreover, the range of effective cell systems for cultivation is limited too. This report highlights the development and comparative evaluation of methods for African swine fever virus (ASFV) growth in porcine bone marrow cell (PBM) and leukocyte (PL) cultures for their efficacy. We showed the possibility of using cell suspension prepared from porcine bone marrow after its 24-hour storage at 4 to 6 °С for ASFV culture. The virus grown in the PBM cell suspension after the abovementioned storage period for porcine bone marrow had expired did not reduce its infectious activity, and accumulated with titers of 7.25 to 7.50 lg TCID 50/cm 3. The conditions for ASF virus culture in hematopoietic cells using stationary devices like KASM (a cassette unit for monolayer culture), a technique for roll-bottle culture in a circular monolayer or in a cell suspension, and biological reactors (fermentors) for a suspension culture were analyzed. When using these culture methods, ASF virus steadily accumulated in the PBM or PL cells with high titers ranging from 7.25 to 8.00 lg HAU 50/cm 3. Nevertheless, ASF virus growth in hematopoietic cell suspensions using automated reactors was found to be the most effective, technologically advanced and productive technique. In this case, the virus titers were as high as 7.50 to 8.00 lg HAU 50/cm 3. Analyzing the results of the studies we concluded that the above methods were more efficient as compared to ASFV growth in a stationary cell culture using flasks, and could be used for large scale cultivation of this biological agent in vaccinology and diagnostics of African swine fever infection.