METHOD FOR CRYOPRESERVATION OF HAEMOPOIETIC UMBILICAL STEM CELLS Russian patent published in 2015 - IPC A01N1/00 A61K35/00 

FIELD: medicine.

SUBSTANCE: cryoprotector is opened in a laminar flow unit; a special syringe of asyringe pump is filled with a cryoprotector; that is followed by introducing a filter solution of the cryoprotector - 55% dimethylsulphoxide with 5% dextran 40 at temperature +4°C in a nucleated cell suspension with haemopoietic stem cells in a cryopackage with the concentrate and mixing mechanically in a mixing apparatus, transferring the system together with the cryoprotector flask into the laminar flow unit; the air is released from the cryopackage and portion of the suspension; the package is sealed and placed into a shrink bag; that is followed by programmed multi-stage freezing, the first stage of which keeping the mixture of the suspension with the stem cells and cryoprotector - a freezing sample - for 10 min at temperature +4°C; the second stage is cooled at a rate of 1°C/min to temperature -12°C; the thirst stage provides cooling at a rate of 20°C/min to temperature -60°C; at the fourth stage, the sample is heated at a rate of 10°C/min to temperature -18°C; at the fifth stage, the sample is cooled at a rate of 1°C/min to -60°C; at the end of the freezing program, the sample is cooled at a rate of 3°C/min to temperature -100°C; after freezing, the sample is placed into a quarantine dewar with liquid nitrogen until infection and bacteriological fungal contamination test results are obtained. After termination of the quarantine shelf life, the sample with haemopoietic stem cells are placed for long-term storage at temperature not exceeding -150°C, into the dewar with liquid nitrogen if observing negative test results. If the infection and bacterial and/or fungal contamination test results are positive, the sample with haemopoietic stem cells are transferred into the dewar with liquid nitrogen for infectious material for long-term storage.

EFFECT: invention enables increasing cell viability in the sample.

3 cl, 4 dwg