METHOD FOR PRODUCING SPHEROIDS OF HEPARG CELLS IN A MEDIUM WITHOUT DIMETHYL SULPHOXIDE Russian patent published in 2018 - IPC C12N5/00 C12N5/74 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology, namely to production of spheroids of HepaRG cells expressing cytochromes P450 3A4, 2B6, 2C19, 2C9. Method comprises culturing HepaRG cells in a complete William's E culture medium supplemented with L-glutamine, 10 % fetal bovine serum, 5 mcg/ml recombinant human insulin, 5x10^-5 M hydrocortisone hemisuccinate, 1 % penicillin/streptomycin, in CO₂-incubator at 37 °C, 95 % air, 5 % CO₂ and relative humidity of 98 %, induction. Method further includes transfer to 96-well plates with low adhesion and a U-shaped bottom to form spheroids and the subsequent incubation of the spheroids in serum-free William's E nutrient medium supplemented with L-glutamine, 1x Insulin-Transferrin-Selenium, 1 mcg/ml bovine serum albumin, 5x10^-5 M hydrocortisone hemisuccinate, 1x interchangeable amino acids, 1 % penicillin or streptomycin at 37 °C, 95 % air, 5 % CO₂ and a relative humidity of 98 %.

EFFECT: invention allows to obtain spheroids of HepaRG cells expressing cytochromes P450 3A4, 2B6, 2C19, 2C9, to study the metabolism and toxicity of pharmaceuticals.

4 cl, 2 dwg, 2 ex