METHOD OF OBTAINING CULTURE OF PACIFIC SALMON CELLS

摘要

FIELD: medicine.;SUBSTANCE: fins of young fish are processed with Hanks' solution, containing 1 mcg/ml amphotericin B and 200 units/ml hentamicin, 200 g are centrifuged for 10 min at 4°C, not less than 5 times. Mechanic crushing of tissue into pieces with size 1 mm in medium DMEM with glucose content not less than 4.5 g/l, 10% of embryonic calf serum, 40 units/ml hentamicin, is carried out. After that, it is incubated in medium with addition of 1 mg/ml of collagenase of type 1 for 60 min at temperature 18-20°C. Collagenase is washed off by double centrigugation, sediment is resuspended with medium DMEM with glucose content not less than 4.5 g/l, 10% of embryonic calf serum, 25 mM Hepes, 40 units/ml hentamicin, 0.25 mcg/ml amphotericin B and 10 ng/ml of human recombinant main fibroblast growth factor. Product is placed into culture vessel for incubation at temperature 18°C with replacement of medium every 3-4 days. When cells achieve monolayer, reinoculation is carried out with application of tripcin or mixture trypsin versen.;EFFECT: method makes it possible to obtain culture of Pacific salmon cells.;3 dwg, 2 ex