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Department of Cryocytology
Research Fellows:

Department was founded in April, 1993. The Head of the Department is Dr. Lyubov A. Babijchuk, DSc (Biol). The principal direction of department scientific activity is the study of molecular mechanisms of structural and functional changes in plasmatic membranes and cytoskeletal proteins of donor and human cord blood erythrocytes, being the base for both cell cryosensitivity and cryoresistance to the low temperature preservation factors.

Head of the Department
Lyubov A. Babijchuk
DSc (Biol)

The investigation of mechanisms for temperature and osmotic erythrocyte stabilisation to factors of low temperature preservation allowed to reveal a determining role in these processes of cell cytoskeleton proteins, which modification resulted in an increase of erythrocyte resistance to the effect of freeze-thawing factors under their cold exposure conditions in PEO-1500 solutions.

The usage of electrophoresis method in PAAG by applying the diamide cross-linked agent allowed to establish the fact, that the erythrocyte incubation with PEO-1500 cryoprotectant at 22C caused the disorder in a spatial localisation and rearrangement of protein fractions of spectrin-actin complex, resulting in cell destabilisation. In contrast the erythrocyte incubation with a cryoprotectant at 0C contributes to stabilisation of cytoskeletal protein structure, preserves their spatial and orientation localisation in a cell and improves the cryopreservation outcomes [1].

The orientation of structural modification of plasmatic membranes and cell cytoskeletal proteins was shown to be determined by the effect of temperature and dehydration level under PEO-1500 non-penetrating cryoprotectant effect on the systems, controlling their structural and functional state. This is related to the changes in intracellular Ca2+ concentration and a level of protein phosphorylation [2].

There was found out the relationship between the character of PEO-1500 binding with erythrocyte membranes at the incubation stage, a change in their volume and shape, modification of cytoskeletal proteins, state of membrane barrier properties and erythrocyte resistance to freeze-thawing [3]. Under conditions of incubations of cells at 0C, when a decrease in cryoprotectant binding with cells is observed, there are occurred structural and functional rearrangements of plasmatic membranes and cytoskeletal proteins, resulting in erythrocyte volume and shape "fixing" and providing in many respects the preservation of membrane barrier properties, that finally contributes to the augmentation of erythrocyte resistance to cryopreservation.

As a result of conducted study of cell stabilisation mechanisms to the effect of a non-penetrating cryoprotectant and low temperatures there were substantiated the new approaches for a targeted modification of structural and functional cell state on the initial stages before cryopreservation with the aim to increase their resistance to freeze-thawing [4].These approaches are in the formation of the definite temperature-osmotic conditions, under which the stabilisation of plasmatic membranes and cytoskeletal proteins occurs. Basing on the principle of "cold stabilisation" the without washing-out way for erythrocyte cryopreservation, allowing in a great extent to increase their resistance to freeze-thawing and to preserve structural and functional cell indices at a quite high level, was developed and patented in Ukraine [5]. This method is more perspective, comparing to the existing ones and can get more wide usage in practical medicine [6].

Nowadays a special attention is paid to the investigation of human cord blood. We investigate the influence of cryoprotectants with various effect mechanisms and cryopreservation process on erythrocytes and other cord blood components [7]. The structural and functional integrity of cord blood cells after cryopreservation has been determined. Cryopreservation method without washing-out for the whole human cord blood preparation to be used in clinical practice has been developed.

23, Pereyaslavskaya str., Kharkov, Ukraine 61016
Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine
Tel: +380 57 373 4143,   373 3807,   373 3039;     Fax: +380 57 373 3084
E-mail: cryo@online.kharkov.ua