MAKEEV BORIS LAVROVICH,Макеев Борис Лаврович,IVOLGIN DMIRIJ ALEKSANDROVICH,Иволгин Дмирий Александрович,SMIRNOVA ANASTASIJA VLADIMIROVNA,Смирнова Анастасия Владимировна,KOROVINA KSENIJA VLADIMIROVNA,К
申请号：
RU2012111734/13
公开号：
RU0002554405C2
申请日:
2012.03.22
申请国别(地区):
RU
年份:
2015
代理人:
摘要：
FIELD: medicine.SUBSTANCE: invention refers to medicine, particularly to haematology. The cryogenic preservation of haematopoietic umbilical cord stem cells is carried out with a cryoprotectant solution - dimethylsulphoxide - added to a suspension of nuclear cells with the haematopoietic stem cells. That is followed by preparing for freezing by cooling the stem cell suspension in a cooling chamber to a temperature of +4°C. The multi-staged freezing of the stem cell suspension involves using a cryoprotectant solution that is 55% dimethylsulphoxide with 5% dextrane 40, which is added into a suspension of a leukocyte concentrate with the stem cells packed in a cryobag. That is followed by mixing mechanically in a mixing apparatus and added with a cryogenic preserving agent at a temperature of +4°C, de-aerating the cryobag with releasing a portion of the suspension, closing the bag, sealing it and placing into a shrink bag and freezing within several stages with computer assistance. At the first stage, the stem cell suspension mixed with the cryoprotectant solution (hereinafter referred to as a sample) is kept for 10 min at a temperature of +4°C, frozen at 1°C/min to a temperature of -12°C, cooled down at 15°C/min to a temperature of -60°C after the sample is unfrozen at 10°C/min, it is cooled down at 1°C/min to -60°C at the end of the freezing programme, the sample is cooled down at 3°C to -100°C. Upon completion of the freezing programme, the sample placed into a cryobox is placed into a quarantine vacuum flask containing liquid nitrogen to determine test results for the presence or absence of infectious agents and bacteriological and mycotic contamination. At the end of the quarantine period, the haematopoietic umbilical cord stem cell sample is transported into the vacuum flask containing liquid nitrogen for long storage at min -150°C with negative test results.EFFECT: invention enables increasing the cells viability.1 tbl, 1 dwgИзобретение относится к области медиц
