METHOD OF SEPARATING FIBROBLASTS, METHOD OF CREATING THEREOF-BASED BIOTRANSPLANT (VERSIONS) AND METHOD OF REGENERATING HUMAN TISSUES (VERSIONS)

摘要

FIELD: chemistry.;SUBSTANCE: invention relates to field of biotechnology and can be applied for regeneration of tissues of mesodermal origin with application of adult organism fibroblasts. Sampling of skin tissues is carried out, obtained bioptate is washed in physiological solution with human albumin, after which it is processed with solution of dispase in culture medium DMEM-LG with glucose content 1 g/l until epidermis is separated from derma. Obtained derma is washed with DMEM-LG and mechanically crushed, derma fragments are distributed around Petri dish and poured with DMEM-LG with foetal bovine serum (FBS), solution of non-essential amino acids (MEM NEAA) and antibiotics. After that, cells are cultivated until fibroblasts fill the entire bottom surface of dish, then they are removed with enzyme, washed, filtered and precipitated by centrifugation. Cell sediment is suspended in growth medium and cultured in flasks in CO₂-incubator until confluent monolayer is obtained, after which cells are removed with enzyme, precipitated and passaged until required therapeutic quantity. Obtained fibroblasts are used for manufacturing transplants for human tissue regeneration.;EFFECT: invention makes it possible to reduce bioptate volume and obtain homogeneous culture of fibroblasts with high degree of adaptation and activity.;29 cl, 8 dwg, 1 tbl, 7 ex