CELL PRODUCT OF INSULIN-PRODUCING MAMMALIAN CELLS AND USE THEREOF FOR DIABETES MELLITUS THERAPY Russian patent published in 2018 - IPC C12N5/00 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology. Invention is a method for producing a cellular product of insulin-producing human cells, comprising producing epithelial progenitor cells from an epithelial tissue biopsy material and their subsequent pancreatic differentiation into cells capable of glucose-dependent secretion of insulin, in which pancreatic cells are differentiated in two stages in a culture medium for eukaryotic cells providing a calcium ion concentration in the range of 0.5 mM to 2.5 mM: at the first stage, cells are cultured for 4–15 days in a culture medium containing at least blood serum – 2–20 vol. %, glutamine – not less than 1 mM, epidermal growth factor – 1–300 ng/ml, transferrin – not less than 0.1 mcg/ml, sodium selenite – 0.1–20 ng/ml, retinoic acid – 0.1–20 mcM, isoproterenol – 0.1–10 mcM; at the second stage, cells are cultured for 4–15 days in a culture medium containing at least blood serum – 2–20 vol. %, glutamine – not less than 1 mM, epidermal growth factor – 1–300 ng/ml, transferrin – not less than 0.1 mcg/ml, sodium selenite – 0.1–20 ng/ml, retinoic acid – 10 nM – 20 mcM, nicotinamide – 1–100 mM, hepatocyte growth factor – 1–300 ng/ml, dexamethasone – 0.01–5 mcM, where the cultivation in both stages is carried out at 37 °C in the presence of 5 % CO₂. Group of inventions includes a cellular product of insulin-producing mammalian cells, as well as a method of diabetes mellitus replacement therapy using a cell product.

EFFECT: invention provides a simplified technology of producing insulin-producing cells, allows producing not less than 70 % of the functionally active insulin-producing cells in the cell culture that has undergone differentiation.

14 cl, 10 tbl, 7 ex, 11 dwg