Роль биологически активных веществ в поддержании плюропотентных свойств ранних зародышей в культуре in vitro
2009
Shejko, I.P. | Gandzha, A.I. | Simonenko, V.P. | Letkevich, L.L. | Rakovich, E.D., National Academy of Sciences. Scientific and Practical Center of Animal Breeding (Belarus) | Motuzko, N.S., Vitebsk State Academy of Veterinary Medicine (Belarus)
Efficiency of cultivation of cow preimplantation embryos and support of pluripotent properties in different nutritive media was analyzed; a method of getting early embryos in vitro for the genetically engineered purposes was developed in the conditions of the Republic of Belarus. Oocyte separation was realized in 3; 3-6; 6-9; and 9-12 hours after ovariectomy for the studying of the influence of storage period of ovaries on the output qualitative embryos. Oocyte separation was realized by bisection with adding of 1% of fetal cattle serum, 10 units/ml of gentamicin and 1 unit/ml of heparin. Oocyte-cumulus complexes maturing took place in CO2-bath with 5% of CO2 in atmosphere of maximum humidity and temperature of 38 deg C in TS-199 (Sigma) media with entering of 25 mM/l of buffer Hepes, 10 unit/ml of gentamicin and biologically active substance (20 % of fetal calf serum, 10 mg/ml of bovine serum albumin and 5 % of estrous cow serum with 10 mg/ml of bovine serum albumin). Research result showed that application of 10 mg/ml of bovine serum albumin into TS-199 media made it possible to increase the yield of cattle embryos at the preimplantation stages in vitro conditions on 19,1%, as well as to increase the fetal calf serum in quantity 15% to the nutritive media volume up to 16,0 %. Application of sodium pyruvate and calcium lactate in the process of cultivation of early cattle embryos obtained in vitro conditions made it possible to increase the yield of preimplantation embryos on 5,4 %. Specifically, the level of transformation of morulas into blastocysts was 44,4 %
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