METHOD OF PRODUCING AND CONCENTRATING microRNA-CONTAINING EXOSOMES OF MULTIPOTENT MESENCHYMAL STROMAL CELLS FOR USE IN COSMETIC AND MEDICINAL AGENTS FOR STIMULATING REGENERATIVE PROCESSES AND SLOWING DOWN AGING PROCESSES
FIELD: medicine.SUBSTANCE: invention refers to medicine, namely biotechnology, and can be used to produce and concentrate micro-RNA-containing exosome multipotent mesenchymal-stromal cells. Method involves production of mesenchymal-stromal cells of umbilical cord, adipose tissue and tooth pulp, subsequent cultivation and collection of culture fluid enriched with exosomes, concentration of exosomes and depletion of large molecular proteins in order to reduce sensitization, freezing or drying of the obtained concentrate for long-term storage. Exosome source used is multipotent mesenchymal stromal cells (MMSC) 1 to 4 passage obtained from tested donors, wherein the cell sources are a dental pulp, adipose tissue, bone marrow, umbilical cord, to accumulate exosomes, MMSC are cultured in medium with low-glucose alpha-MEM or DMEM, 5 mM L-alanyl-glutamine and activated platelet plasma urea-cord plasma (UCPRP), culturing the cells in the medium described above is carried out up to 80–100 % of the monolayer of the culture, wherein culturing can be static or flowing: in static cultivation, cells are placed in a culture dish such as a bottle and a petri dish. When monolayer reaches 80–100 %, the culture fluid is changed every 24 hours, the collected culture medium is stored for exosome release, wherein from one culture is 5–7 removable medium, and in flow cultivation cells are placed in a flow type incubator on cellular carriers, such as microparticles, hollow-fiber structures, when cells reach the monolayer, the culture medium is completely changed to a new one, after which standard cultivation is performed for 7 days. When obtaining medium enriched with exosomes, its concentration is carried out with removal of protein fraction of more than 100 kDa, using a system of tangential filtration to remove coarse particles and concentration: first, the culture medium is centrifuged for 30 minutes at 2000g or filtered by 1–10 mcm filter to remove large cell debris, then the supernatan
