METHOD OF PRODUCTION OF PRIMARY CELL CULTURE OF KIDNEYS OF NEWBORN RABBITS FOR REPRODUCTION OF VIRUSES

摘要

FIELD: biotechnology.;SUBSTANCE: method comprises euthanasia of infant rabbits by decapitation, removing paws, tails, removing skins, opening the abdomen cavity at the level of the lumbar vertebrae, kidney extraction, removal of the organ capsule, shredding into pieces with the size of 1-3 mm. The donors of the primary cell culture the newborn rabbits 1-2 days old are used. The pieces of tissue are washed from residual blood with saline solution of Hanks, subjected to disaggregation in 0.25% trypsin solution by preliminary incubation of the pieces at 37°C for 30-40 minutes, the cells are pelleted by centrifugation at 1000-1500 rev/min for 10 minutes, the supernatant is drained off, and the pellet is resuspended in the growth medium consisting of medium Igla MEM, 0.5% solution of lactalbumin (1:1) with the pH 7.0-7.2 with 10% serum of adult cattle or foetuses of cows, the cell concentration is adjusted to 5-7·105 cells/cm³, then antibiotic ciprofloxacin at 100 U/ml is added. At that the sensitivity of primarily trypsinised cultures of kidney cells of newborn rabbits to respiratory-enteric viruses of cattle, parvoviruses, herpesviruses and parainfluenza-3 by determining the viral titers in lg 50/ml TDC is determined.;EFFECT: method enables to enhance the sensitivity to viruses of animals, to improve the biological activity of the cultured cells, and to increase the yield of viral material.;3 tbl, 3 ex