CELL CULTURE AND BIOTRANSPLANT FOR BONE TISSUE REGENERATION ON ITS BASIS Russian patent published in 2018 - IPC C12N5/00 

FIELD: medicine; biotechnology.

SUBSTANCE: invention relates to the field of biotechnology, in particular to the production of multipotent mesenchymal stromal cells cultured as spheroids and induced in the angiogenic direction. Method includes isolating a donor tissue fragment containing multipotent mesenchymal stromal cells, transferring a donor tissue fragment to a medium containing culture medium, an amino acid and an antibiotic, mechanical grinding of a fragment of donor tissue with obtaining a homogenate, incubating the obtained homogenate in a solution of proteolytic enzymes at 37 °C until receiving the cell suspension. Next, add a nutrient medium containing culture medium, amino acid, antibiotic, serum; the cell suspension is centrifuged until the cells are completely precipitated, the supernatant is removed, the isolated cells are resuspended in a nutrient medium containing culture medium, an amino acid, an antibiotic, blood serum, cultivation of selected cells in monolayer culture at 37 °C in an environment of 5 % CO₂ at least 2 passages, monolayer disaggregation, cultivation of cells of at least 2^nd passage in non-adhesive conditions in concentration from 1 × 10⁶ cells/ml to 5 × 10⁶ cells/ml in a nutrient medium containing culture medium, amino acid, antibiotic, blood serum, angiogenesis induction factor to produce a cell culture containing multipotent mesenchymal stromal cells cultured as spheroids.

EFFECT: invention allows to obtain multipotent mesenchymal stromal cells cultured as spheroids and induced in the angiogenic direction.

25 cl, 3 dwg, 3 ex