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
Title | Year | Author | Number |
---|---|---|---|
METHOD FOR OBTAINING LINING CELLS FROM THE OLFACTORY LINING OF MAMMALS FOR THE TREATMENT OF SPINAL CORD INJURIES | 2017 |
|
RU2676142C2 |
PRODUCTION OF TERMINAL-DIFFERENTIATED DOFAMINNERGIC NEURONES FROM EMBRIONIC HUMAN FOUNDER CELLS | 2004 |
|
RU2345133C2 |
PRODUCTION OF GABA-ergic NEURONS in vitro OF EMBRYO STEM CELLS AND APPLICATION THEREOF IN TREATING NEURAL DISORDERS | 2004 |
|
RU2380410C2 |
METHOD FOR PRODUCING NEURONAL MATRIX | 2010 |
|
RU2431666C1 |
TRANSPLANTATION OF NERVE CELLS FOR TREATMENT OF NEURODEGENERATIVE STATES | 2005 |
|
RU2434636C2 |
METHOD OF SPINAL TISSUE ENGINEERING FOLLOWING ANATOMICAL RUPTURE | 2012 |
|
RU2489176C1 |
BIOMEDICAL CELL PRODUCT FOR TREATING NERVOUS DISEASES AND MENTAL DISORDERS, METHOD FOR PRODUCTION THEREOF AND USE THEREOF | 2018 |
|
RU2720002C1 |
METHOD FOR TREATING TRAUMATIC SPINAL LESIONS | 2000 |
|
RU2195941C2 |
AGENT FOR TREATMENT OF SPINAL CORD AND BRAIN DAMAGES | 2004 |
|
RU2290939C2 |
EVALUATION METHOD OF MORPHOFUNCTIONAL STATE OF INDUCED PLURIPOTENT STEM CELLS OF PARKINSONIAN PATIENTS, WHICH ARE DIFFERENTIATED TO DOPAMINERGIC NEURONS | 2012 |
|
RU2501853C1 |
Authors
Dates
2021-06-07—Published
2020-07-23—Filed