METHOD OF VITRIFICATION OF OVARIAL TISSUE Russian patent published in 2019 - IPC C12N5/07 A01N1/02 A61K35/54 

FIELD: medicine.

SUBSTANCE: invention relates to the field of medicine, namely to gynecological and reproductive medicine, and is intended to preserve the reproductive function of women with cancer who wish to have children in the future. Method of vitrification of ovarian tissue includes the preparation of ovarian tissue samples, their two-stage saturation with cryoprotectants in a medium containing 7.5 % DMSO solution and 7.5 % EG, 0.5 M sucrose, while tissue fragments are stored in sterile cryovials with liquid nitrogen. Preparation of ovarian tissue samples for vitrification is carried out by removing the cortical layer from the brain layer of the ovary. Thickness of the cortical layer is set to not more than 1 mm and divided into fragments of 2–5 mm × 2–5 mm at +4 °C. Then the tissue fragments are saturated with cryoprotectant solutions in two stages. At the first stage, tissue fragments are impregnated for up to 20 minutes in a vitrification medium containing inorganic salts – 10.5 g/l, amino acids – 1.0 g/l, vitamins – 0.10 g/l, glucose – 1.0 g/l, penicillin-streptomycin (100×) 10 ml/l and DMSO cryoprotectors – 7.5 vol.%, EG – 7.5 vol.%, with the addition of fetal calf serum up to 20 vol.%. At the second stage, the tissue fragments are kept for 10 minutes in a vitrification medium containing inorganic salts – 10.0 g/l, amino acids – 1.0 g/l, vitamins – 0.10 g/l, glucose – 1.0 g/l, penicillin-streptomycin (100×) 10 ml/l and DMSO cryoprotectors – 17.0 to 20.0 vol.% and EG – 10.0 to 13.0 vol.%, sucrose – 0.5–0.6 M. Prepared fragments of the cortical layer of the ovarian tissue are placed in sterile labeled cryovials with liquid nitrogen for storage at -196 °C.

EFFECT: use of the invention allows to reduce the toxic effect and ensure high survival of the follicles and cellular components of the stroma.

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