Work

Series of scientific works consists of 7 scientific articles published over 4 years. This is a study of morphological and functional characteristics of newborn piglets adrenal cell cultures, as well as their changes during cryopreservation, depending on the initial state of the biological material to be frozen. The influence of the enzymatic preparation and culturing conditions on the morphology and hormonal activity of newborn piglets adrenal cells during culturing in vitro was investigated. The paper presents data of immunocytochemistry, radioimmunoassay, biochemical research methods, optical and fluorescence microscopy, morphometric analysis of cell cultures before and after cryopreservation, as well as under their culturing in different conditions. It is shown that when cells are grown in a medium containing 10% FCS the cytosphere formation and neuron migration were observed. The cells migrating from cytospheres express β-III-tubulin, but do not express the S-100. The relevance of the problem and the necessity of cryopreservation experiments were described in detail. The results of primary cell culture and tissue fragments cryopreservation were described. It was shown that after primary cell cultures cryopreservation at a cooling rate of 1 deg / min in media containing 5-15% DMSO with or without FCS (10 or 25%), cortisol secretion was retained, and the formation of a monolayer, cytospheres and neuron cells were observed. It was found that primary cell culture may be obtained from tissue fragments after their cryopreservation under 10% DMSO at a cooling rate of 0.3 ° C / min. In such cultures cytospheres were formed and β-III-tubulin positive cells of neuronal morphology were migrated. Cryopreservation at a cooling rate of more than 1 ° C / min does not provide a monolayer culture, as the cells lose their ability to spreading. These research results were presented in 7 articles in specialized journals, including 1 - in foreign editions; and 5 abstracts at conferences, congresses and symposia.