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Study of spheroidogenesis and plasticity of adrenal cells during in vitro culturing and development of methods for their long-term storage


Presented Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine

Ph.D. Sidorenko O.S.

The paper presents data from immunocytochemical, radioimmunological, biochemical methods of research, light and fluorescence microscopy, morphometric analysis of cell cultures before and after cryopreservation, as well as during their cultivation in different conditions. It has been shown that when culturing cells in a medium containing 10% fetal calf serum (FCS), the formation of spheroids and the migration of neuronal cells from them are observed. Cells migrating from spheroids express β-III-tubulin, but do not express S-100.

It is shown that after cryopreservation of primary cell cultures with a cooling rate of 1 deg / min in media containing 5-15% dimethyl sulfoxide (DMSO) without FCS or with the addition of FCS (10 or 25%), cortisol secretion was preserved, the formation of monolayer and spheroids, as well as neuron-like cells appeared during reculture. It was found that cryopreservation of tissue fragments under the protection of 10% DMSO with a cooling rate of 0.3 deg / min and subsequent enzymatic treatment can allow to obtain primary cell culture, in which spheroids and β-III-tubulin positive cells of neuronal morphology are formed. Cryopreservation with cooling rates greater than 1 deg / min does not allow to obtain a monolayer culture, because the cells lose the spreading ability.

Number of publications: 13 scientific articles, including 5 articles in English-language journals with an impact factor, as well as one utility model patent. The total number of references to the author's publications is 22 (according to Google Scholar) and 5 (according to Scopus), the h-index is 3 (according to Google Scholar) and 2 (according to Scopus).