METHOD FOR OBTAINING ADHESIVE CULTURE OF NEURAL STEM/PROGENITOR CELLS OF OLFACTORY NOSE OF MAMMALS FOR TREATMENT OF SPINAL CORD INJURIES

摘要

FIELD: biotechnology and medicine. ;SUBSTANCE: invention relates to the field of biotechnology and medicine. Disclosed is a method of obtaining an adhesive culture of neural stem / progenitor cells from the olfactory lining of a mammalian nose for the treatment of spinal cord injuries. It includes obtaining a tissue sample from the olfactory lining of a mammal's nose, followed by washing and incubating for 1 hour at 37°C with 3.6 units/ml of dispase II, then the tissue of the olfactory epithelium is mechanically separated from its own plate in DMEM:F12 (1:1) medium with antibiotics and incubated with a solution of 500 U/ml collagenase for 10 min at 37°C, the enzyme is inactivated with medium with serum, resuspended, centrifuged for 2 min at 900 g, cells are placed on plastic and cultured for 5 days in DMEM:F12 medium (1:1) containing 10% FBS, 2 mM L-glutamine and antibiotics, the medium is replaced every two days, formed by the monolayer of olfactory epithelium cells is removed with 0.25% trypsin solution, placed on plastic coated with 50 mcg/ml fibronectin, cultured for 1-2 days in neurobasal medium containing 2 mM L-glut amine, 1% ITS, 2% B27, 100 mcg/ml streptomycin, 100 U/ml penicillin, 60 mcg/ml gentamicin, 50 ng/ml bFGF, 50 ng/ml EGF, the resulting cells are characterized by markers nestin and βIII -tubulin.;EFFECT: obtaining an adhesive culture enriched in neural stem / progenitor cells with increased survival in an amount sufficient for further transplantation.;4 cl, 14 dwg, 5 ex