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Impact of preserving agents upon on intracellular meтaboliс changes during cryoconservation of human bone marrow

UDC 547.963

Impact of preserving agents upon on intracellular meтaboliс changes during cryoconservation of human bone marrow

Vashchenko V. I., Сhuklovin A. B., Petrenko G. I., Vilyaninov V. N., Bagautdinov Sh. M.

Abstract
Bone marrow samples (autologous and donor cells) harvested for transplantation were studied in present work. Typing of lactate dehydrogense (LDH) isoenzymes was performed by means of the SUPER CELLO-5 device (HOSPITEX, Switzerland). We have shown that, the thawed cells exhibited a shift of the LDH isozyme profile from M-forms towards N-forms, thus presuming a fast restoration of cellular metabolism. Changes of the LDH isoenzyme spectrum detected under the influence of ultralow temperatures may reflect an intracellular rearrangement of metabolic processes and intracellular redistribution of LDH isoforms, being an inportant element of cell adaptation to the temperature shifts. First of all, these biochemical changes are in accordance with considerable reduction of ATP contents in thawed cells from the bone marrow when using PEО-400 or PVP as cryoprotectants (resp., 1.5- and 1.8-fold decrease in ATP). Secondly, synthesis of pyruvic acid proved to be sufficiently increased in a suspension of the thawed cells. Thirdly, intactness of the bone marrow cells if concerved with polyvinylpyrrolidone cryoprotectant was considerably higher than when applying the polyethylene oxide (PEO-40) (resp., 85% with PVP versus 57.2% for PEO). Forthly, structural changes of the supercoiled DNA in the cell samples did well correlated (r = 0.92; p < 0.01) with changes in АTP and pyruvate quantities in their extracellular space.

Keywords: cryopreservation, isoenzymes lactate dehydrogenase, supercoiled DNA, biochemisry changes, bone marrow cells, dimetylacetamid, polyvinylpyrrolidone, polyetilenoxid 400

All references

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