METHOD OF HUMAN ROTAVIRUS ADAPTATION TO GROWTH ON TRANSPLANTATED ANIMAL CELL CULTURES Russian patent published in 2019 - IPC C12N7/00 

FIELD: biotechnology.

SUBSTANCE: invention relates to the field of biotechnology. Invention represents a method of adapting group A human rotavirus to growth on transplantable cell cultures of animals, consisting in the fact that adaptation and subsequent cultivation of human rotavirus group A is carried out by successive passages on a mixed culture consisting of epithelioid and fibroblast cell types at ratio of 1:1; epithelioid cells are represented by a transplanted pig embryo kidney cell culture (SPEV), a fibroblast type – a transplanted rat fibroblast culture (RF) grown separately on medium 199; obtained monolayer of cells of both types is removed from glass with a mixture of a solution of Versen and 0.25 % solution of trypsin in ratio of 5:1, diluted with medium 199 in 4 times, mixed at ratio of cell cultures 1:1, added 10 % of heated whey of bovine animals, maintained in thermostat at +37 °C for 2 days; washing 3-fold monolayer of cell mixture from growth medium with Hanks solution, adding 0.5 ml of trypsin-treated (20 mcg/ml) 10 % of faecal suspension containing human rotavirus, and contacting for 10 min; adding support medium 199 without whey to initial volume and holding in thermostat at +37 °C for 10 days; frozen at -18–20 °C for 5 hours with subsequent thawing at 37 °C, repeating the procedure three times; subsequent passages are carried out according to the same scheme using an inoculum of the previous passage.

EFFECT: invention enables producing group A stable rotavirus strains stably growing on transplantable cell cultures with viral particle concentration of not less than 1⋅10⁹ in 1 ml of culture fluid, having a typical morphology of rotaviral particles and double-stranded segmented RNA.

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