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Registration Number

0221U101231, 0119U101733 , R & D reports

Title

Development of a breast cancer cell screening system based on the 3D model of tumor spheroids enriched with cancer stem cells

Head

Sydorenko Mykhailo V.,

Registration Date

18-01-2021

Organization

Department for special training National academy of science of Ukraine at Kiev University

Description

The object of the study is multicellular tumor spheroids of breast cancer cells of the MCF-7 line, enriched with cancer stem cells (CSCs). The purpose of the study is to assay a cell culture system of multicellular tumor spheroids enriched with cancer stem cells (eMCTS) to determine the effect of anticancer drugs. Research methods of the study are cultivation of adhesive cells and multicellular tumor spheroids (MCTS), immunohistochemical, morphometric, spectrophotometric, cell survival analysis (MTT test), counting of the cells in Goryaev's chamber and flow cytometry. To estimate the percentage of stem cell phenotype of tumor cells in MCTS culture after CSCs enrichment, a comparison of markers characteristic of breast cancer stem cells was performed. Thus, after long-term cell cultivation in monolayer monolayer and spheroidal cultures of the MCF-7 line, increased expression of intracellular marker nestin and surface marker CD133 was detected, as well as increased aldehyde dehydrogenase activity by eMCTS cells, compared with adhesive cells and MCTS. Decreased survival and cell aggregates after 48 hours of incubation with salinomycin, a promising drug aimed at eliminating CSCs populations, have been shown compared to control samples. After enrichment of MCTS culture by CSCs, salinomycin inhibited the formation of cell aggregates and had a cytotoxic effect on cell populations of this culture. Interferon alpha, as a factor of the cell microenvironment, also inhibited the growth of tumor cells of the MCF-7 line, promoted cell differentiation and dose-dependently affected the breakdown of MCTS and the formation of a large number of small cell aggregates and single cells. Soluble factors from mesenchymal stromal cells, which are elements of the tumor stroma, in contrast, increased the survival of tumor cells and the formation of larger spheroids in a culture enriched by CSCs. Therefore, the method of culturing eMCTS is a way to maintain the population of CSCs in culture i 

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Dzubenko Natalya V.
Kuznyetsova Galyna M
Perepelytsina Olena M
Sidorenko Mihaylo V.