METHOD FOR OBTAINING PREPARATION FOR STIMULATION OF REGENERATION BASED ON PRODUCTS OF SECRETION OF HUMAN MULTIPOTENT MESENCHYMAL STROMAL CELLS

摘要

FIELD: medicine.;SUBSTANCE: group of inventions relates to medicine and can be used for obtaining biomaterials for stimulation of tissue regeneration after injury. Method of preparation obtaining includes cultivation of human mesenchymal stromal cells of adipose tissue (MSC AT) to 4-5 passage in medium, supporting growth of non-differentiated human mesenchymal cells. Washing of cells with buffer solution, conditioning MSC AT in serum-free and deprived of animal-derived products growth medium, supporting cell viability not lower than 79% and suitable for therapeutic application, are performed. After sampling cultivation medium, containing products of human MSC AT secretion, remnants of cells are removed from it with purification from low-molecular components with the following lyophilisation of purified cultivation medium. Preparation contains products of human MSC, including key growth factors: VEGF in concentration not lower than 200 pkg/ml, HGF in concentration not lower than150 pkg/ml, FGF basic in concentration not lower than 0.29 pkg/ml, angiopoietin-1 in concentration not lower than145 pkg/ml, PEDF in concentration not lower than 500 pkg/ml, determined by method of enzyme immunoassay with reduction of lyophilisate in sterile physiological solution. Preparation efficiency is provided due to selected content in medium of key growth factors, required for realisation of regenerative MSC action, namely vessel endothelium growth factor (VEGF), hepatocyte growth factor (HGF), basic fibroplast growth factor (bFGF) and angiopoietin of type1 (Angpt-1), as well as antiangiogenic factor, obtained from pigment epithelium cells (PEDF).;EFFECT: application of the obtained preparation makes it possible to stimulate tissue regeneration in efficient way due to complex balanced action of growth factors and other biologically active molecules, secreted by human mesenchymal stem/stromal cells (MSC) into culture medium, which does not contain xenogenic components.;12 cl, 9 ex, 6 dwg