METHOD FOR CRYOPRESERVATION OF BIOLOGICAL OBJECTS WITH SIMULTANEOUS HOMOGENEOUS NUCLEATION OF CRYSTALS OF ICE AND XENON CLATHRATE

摘要

FIELD: technological processes.;SUBSTANCE: invention relates to cryopreservation of biological objects, such as cells, tissues, organs. Method for cryopreservation of a bioobject by a combination of cooling and pressure by a clathrate-forming inert gas in a closed volume comprises a step of adding to the initial solution, in which a bioobject, a milled xenon clathrate, prepared based on a solution identical to the initial one and at least one cryoprotector, is placed. At the second stage, the pressure of xenon in the chamber volume is increased to 0.5–3 atm, and then the biological object is cooled to temperatures of -115 °C to -130 °C. After storage, working capacity of the bioobject is restored by fast heating to temperatures from +5 to +20 °C, with subsequent decompression of chamber during 2 to 20 minutes. Further, the biological object is washed from the cryoprotector by multiple medium substitution and the biological object is slowly heated at rate of 0.5–2.0 °C/min to the physiological temperature of the bioobject, after which the biological object is removed from the chamber and placed in a fresh medium without a cryoprotector.;EFFECT: disclosed is a method for cryopreservation of biological objects with simultaneous homogeneous nucleation of crystals of ice and xenon clathrate.;5 cl, 4 dwg