METHOD FOR OBTAINING OF BIOSAFE CULTURE OF MESENCHIMAL STEM CELLS FROM HUMAN CHORIONIC VILLAE Russian patent published in 2018 - IPC C12N5/00 C12N5/78 

FIELD: biotechnology.

SUBSTANCE: method includes mesenchymal stem cells culture selection from human placenta after caesarean section from chorion villae, tissue disaggregation by a solution containing 0.2% of type 2 collagenase and 0.18% of type 4 collagenase prepared on DMEM\F12 medium at 1:1 ratio, and incubation at 37°C for 30 min. Further, the separated cells are collected, centrifuged at 800 rpm for 15 min, obtaining cell sediment, which in resuspended in the DMEM\F12 growth medium, containing 5% of fetal calf serum, 100 u/ml of penicillin, 100 g/ml of streptomycin, 2 mM of glutamine, 1 mM of sodium pyruvate, the cell sediment is resuspended in the DMEM\F12 growth medium (ratio is 1:1), transferred to culture flasks and cultured in an atmosphere containing 5% of CO₂ till 3\4 of monolayer is formed, changing the growth medium 3 times a week, and when 3\4 of monolayer is reached, cells are reinoculated in a ratio of 1:2, and after 1 passage a part of cells is use to detect AIDS, hepatitis B and C and mycoplasma viruses in the culture by PCR, and in case of positive analysis on these infection the culture must be rejected.

EFFECT: invention allows to obtain a biosafe culture not infected by AIDS, hepatitis B and C, and mycoplasma viruses.

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