Dudetska G. Influence of cryopreservation factors on zonally differentiated populations of animal adrenocytes

The object of research is the influence of cryopreservation conditions on the morphofunctional properties of the total suspension of cells, cortical cells, medullary cells, cell fractions of the total cell suspension of adrenal glands (AG). The goal of study was to analyze the effect of the composition of a cryoprotective medium containing different concentrations of dimethyl sulfoxide (DMSO), the temperature of DMSO saturation, and different cooling rates on the morphological and functional characteristics of the total cell suspension, fractions of the total cell suspension and cells of the adrenal cortex and medulla. Research methods: the method of cell enzymatic isolation, cultivation, cryopreservation, histochemical methods, light and fluorescence microscopy, morphometric analysis, volumetric method, methods of statistical analysis. For the first time, separation of the total cell suspension of AG in a ficoll density gradient was carried out, which made it possible to isolate cell fractions, each of which was enriched with a zone-specific cell type, that was confirmed by histochemical methods. It was found that saturation in DMSO solutions at a temperature of 4 °C is the most favorable for the total cell suspension and for the adrenal cortex cell suspension. For the first time, the permeability coefficients of the membranes of the cells of the adrenal cortex and medulla for water and DMSO molecules as well as the values of the activation energies of the processes of transfer of these substances through the cell membranes were determined. It was found that low cooling rates (1, 5, 10 °C/min) make it possible to preserve the greatest amount of the total suspension of AG cells and adrenal cortical cells. It was established for the first time that cryopreservation of the total cell suspension of AG at a cooling rate of 10 °C/min in the presence of 7% DMSO (saturation at 4 °C) provided the maximum number of preserved steroid-producing cells, which retained morphological and functional properties similar to the native cell culture after warming. Field of Application: cryobiology, biology, medicine, biotechnology, veterinary, pharmacy.