Maliukina M. Sensitivity of transmembrane potentials of isolated hepatocytes to agonist-dependent regulation on the different stages of cryopreservation.

The research object: influence of different stages of cryopreservation on the dynamics of regulation of cytoplasmic and mitochondrial potentials under conditions of activation of cells by alpha-agonist. The research aim:estimation of functional integrity of signal systems of hepatocytes and their functional state on the different stages of cryopreservation by means of the test-system based on the microfluorimetric determination of changes of transmembrane potentials under agonist action. Research methods: microfluorimetry, microspectrofluorimetry, luminescence spectroscopy. It was determined, that changes of hepatocyte plasmatic membrane transmembrane potential in response to phenylephrine have phase character with two phases of hyperpolarisation that are regulated by different mechanisms. This character of plasmatic membrane potential induced oscillations preserves at estimation by means of oppositely charged DiOC2 and SqSC4 dyes. Changes of cell potential were accompanied by rise of mitochondrial potential that was determined by increase of JC-1 aggregates fluorescence in hepatocytes. It was determined that mitochondrial response dynamics peculiarities provide the information about integrity and acting of signal transfer systems in cells. The possibility of this approach use for estimation of functional state of the cells, which were cryopreserved was shown. Fluorescent estimation of potential agonist-induced oscillations allows to determine the latent disturbances in intracellular signal transfer systems at cryopreservation and to compare the effectiveness of different protector agents for prevention and correction of these disturbances.