METHOD FOR PREPARATION OF MULTIPOTENT STROMAL CELLS FROM CRYO-FROZEN FETOPLACENTAL COMPLEX TISSUES Russian patent published in 2017 - IPC A01N1/02 

FIELD: medicine.

SUBSTANCE: invention refers to cryobiology and medicine. For multipotent stromal cells (MSCs) reception from frozen fetoplacental complex tissue (umbilical cord, fetal part of the placenta, amnion), the umbilical cord and placenta, obtained during cesarean section, are transported to the culture laboratory within maximum 24 hours, where they are cut into pieces with thickness of max. 10 mm and kept in cryoprotective medium based on the autoplasma obtained from umbilical cord blood or phosphate buffered saline with pH 7.4, supplemented with sampled human albumin to 15 g/l, containing 1.5 mol/l of propanediol and 0.1 mole/l of sucrose, for 20-25 minutes. Further, the fragments of tissue are subjected to gradual cooling: initial room temperature lowering down to +4°C ata rate of 10°C/min, cooling down to -6°C at a rate of 2°C/min, initiation of crystallization at -6°C, step-by-step cooling down to a temperature of -30°C at a rate of 0.3°C/min, immersion in liquid nitrogen with a temperature of -196ºC for prolonged cryo-storage. For defrosting, the tubes placed in a water bath heated to +37°C, and then washed free of cryoprotectant heated by full cultural medium free from xenogenic components and heated to +37°C. Then, epithelium and blood vessels are removed from the umbilical cord fragments, chorionic villi and chorionic trophoblast are removed from the placenta fragments, amniotic epithelium is removed from the amnion fragments, and the remaining tissue (Wharton's jelly of the umbilical cord, placenta fetal portion stroma and amnion stroma) are milled by means of surgical instruments in a small volume of culture medium to obtain explants volume of 1-2 mm³, that is transferred to culture vessels to produce a primary cell culture. Further growth and characterization of MSCs are performed using standard culture techniques.

EFFECT: invention provides MSCs cultures obtained from frozen fetoplacental tissue by maintaining the viability of the cellular stromal component in cryopreservation of tissue without application of of xenogenic and toxic media components.

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