METHOD OF PRODUCTION OF SHEEP EMBRYOS in vitro

摘要

FIELD: biotechnology.;SUBSTANCE: method comprises extracting the oocytes-cumulus complexes from ovaries, cultivating them to the stage of metaphase II, co-cultivating of oocytes and sperm cells, cultivating the impregnated ootids. And additionally the follicle-stimulating and luteinising hormone is administered in the form of the combined preparation Menopur, estradiol, heparin, caffeine and antibiotic. The cultivating is carried out in the base medium. Extraction of the oocytes-cumulus complexes from ovaries is carried out from live sheep under anaesthesia by puncturing ovarian follicles during laparotomy. Superovulation stimulation is preliminary carried out. In the maturation medium the preparation Menopur is added, the concentration of both components that are part of the preparation, is 0.075 IU/ml, and estradiol is added to the maturation medium in the concentration of 10 mcg/ml. For impregnation the newly obtained sperm of tups is used. For tup sperm capacitation and to the maturation medium the combination of heparin is added in an amount of 5 units/ml and caffeine in an amount of 0.2 mg/ml. The cultivation and impregnation is carried out in four well Petri dishes in 500 mcl nutrient medium under 200 mcl paraffin oil.;EFFECT: use of the method enables to increase the proportion of output of oocytes.;3 cl, 11 dwg, 3 ex