FIELD: agriculture.
SUBSTANCE: invention refers to medicine - transfusiology. Nuclear cells with hematopoietic stem cells (HSC) of cryoprotector - 55% of dimethyl sulfoxide solution with 5% dextran 40 arranged in a cryobag are added into a suspension. Mechanically mixed in a mixing device at +4°C, the cryobag is sealed and put into a shrink-wrap bag. Programmed freezing is carried out, at the first stage of which a sample is maintained for 10 min at +4°C, then cooled with the speed of 1°C/min down to the temperature of -12°C, then cooled down with the speed of 15°C/min down to the temperature of -60°C. After the sample thawing with the speed of 15°C/min down to the temperature of -18°C, it is cooled with the speed of 1°C/min down to -60°C . At the end of the freezing program the sample is cooled with the speed of 3°C/min down to -100°C. On completion of freezing the sample is put into a quarantine Dewar vessel with liquid nitrogen until results of tests for infection availability are ready. On completion of a quarantine period of storage, the sample is transferred for long-term storage at the temperature of at least -150°C into a Dewar vessel with liquid nitrogen provided the tests results are negative. In case the tests results are positive, the sample with HSC are transferred to a Dewar vessel for infection material for long-term storage.
EFFECT: invention makes it possible to increase HSC sterility during freezing, integrity and their viability.
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Authors
Dates
2011-04-20—Published
2009-12-11—Filed