FIELD: biotechnology.
SUBSTANCE: method of culturing tumour stem cells of glioblastoma is disclosed. The method comprises selecting such cells, inoculation and incubation in the culture vessel. At least 0.5 g glioblastoma material is selected, which is crushed to pieces no larger than 5x5x5 mm, which are not allowed to dry. Then, the tumour stem cells are isolated from the selected material and they are precipitated. Then they are mixed with the medium for culturing mammalian cells, the cells are resuspended and precipitated into culture flasks. Culturing is carried out to form a monolayer. Then enzymatic dissociation is carried out for 10 minutes at 37°C by 0.05% trypsin-ethylenediaminetetraacetic acid added in a ratio of 1:4, centrifugation for 3 min at 1200 rev/min. Then fresh medium is added, resuspended, re-cultured using the cell suspensions containing no less than 85-95% viable cells, and treated with a fluorescent marker fluorescent under ultraviolet radiation. The invention can be used in laboratory pre-clinical molecular-biological evaluation of the therapeutic efficacy of anticancer preparation obtained by targeted modification of transcriptomic profile of hematopoietic stem and progenitor cells (SC and PC) used for high-tech complex treatment of preferably malignant glial brain tumours.
EFFECT: improvement of the method.
4 cl, 2 ex
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Authors
Dates
2016-01-20—Published
2014-12-05—Filed