METHOD OF PRODUCING CATTLE EMBRYOS IN VITRO Russian patent published in 2024 - IPC A01K67/02 A61D19/04 

FIELD: biotechnology; veterinary science.

SUBSTANCE: disclosed is a method for producing cattle embryos in vitro, comprising transporting cow ovaries from a slaughterhouse in a saline solution containing heparin, gentamycin and bovine serum albumin, extraction of oocytes from ovaries using an aspirator with a trapping vessel FTA-2i with 18 G needle into IVF1 "Aspiration" medium containing antibiotics and phenol red, then oocyte-cumulus complexes are washed in IVF1 "Universal" medium containing antibiotics and phenol red, and cultured in IVF1 "Universal" medium for 21–24 hours in incubator at 38.6–38.9 °C, content of CO₂ 6.2–6.8% and uncontrolled humidity without oil coating. Cryopreserved sperm is thawed in water at 37 °C, then, ejaculate is subjected to gradient fractionation with the use of ready solutions “Perselect” 90% and “Perselect” 45%. Co-cultivation of mature oocytes and sperm cells is carried out in four-well plates in IVF1 "Universal" medium for 16–20 hours in incubator at 38.6–38.9 °C, content of CO₂ 6.2–6.8% and uncontrolled humidity without oil coating; fertilized eggs are cultured in IVF PRO “Droblenie” medium containing antibiotics and phenol red without oil coating; at stage of 4–8 blastomeres for subsequent cultivation, the embryos are transferred into the IVF PRO "Blastnaya" medium containing antibiotics and phenol red without oil coating, and incubated at 38.6–38.9 °C, content of CO₂ 6.2–6.8% and uncontrolled humidity without oil coating to the blastocyst stage.

EFFECT: invention provides a wider range of methods for producing cattle embryos in vitro outside an animal’s body in a non-core laboratory, not requiring preparation of working solutions for each stage of cultivation, for research purposes and livestock breeding.

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