METHOD FOR PRODUCING IN VITRO CATTLE EMBRYOS Russian patent published in 2024 - IPC C12N5/10 A01K67/02 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology, namely to cell engineering, and can be used to create a system for producing cattle embryos in vitro. Invention represents a method of producing cattle embryos in vitro. Method includes ovaries sampling from cows after slaughter, obtaining oocyte-cumulus complexes, their maturation in a commercial medium for in vitro development of human embryos with addition of follicle-stimulating hormone and human chorionic gonadotropin, washing of mature in vitro oocytes, processing of bovine sperm cells, fertilization by joint incubation of oocytes with sperm cells in a commercial medium for human embryos in vitro for 24 hours at temperature of 38.5 °C using heparin as a capacitating agent and cultivation of fertilized oocytes in a commercial medium for human embryos at temperature of 38.5 °C. As a commercial medium for human embryos, "Continuous Single Culture Complete" medium is used, which in fresh form at the stages of maturation, fertilization and in vitro cultivation of cattle embryos each time is coated with mineral oil for cell cultures "Sage", at a carbon dioxide level of 6.5 vol.% and oxygen 5.0 vol.%, wherein sperm cells are processed by centrifugation for 15 minutes, at 15,000 rpm under conditions of room temperature and in density gradient: 3 ml of 80% Percoll, and the sperm sediment after centrifugation for 10 minutes at 8,000 rpm is washed with a buffer medium containing 3 IU of heparin, and motile sperm cells are introduced into the fertilization medium with their concentration of 1.0–2.0×10⁶ in 1 ml of this medium and at rate of 10 oocyte-cumulus complexes per 50 mcl of medium, then, 16–18 hours after fertilization, the complexes are cleaned from cumulus cells and sperm cells, transferred into a medium and cultured for 170–180 hours to the blastocyst stage.

EFFECT: invention makes it possible to increase potential of embryo development without variability from batch to batch and to simplify process of cattle embryos production.

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