Livestock Studies 2007, Vol 47, Num, 2     (Pages: 001-005)

The Vitrification of In Vitro Fertilized Bovine Embryos after 7 Days Cultured

Sedat Hamdi KIZIL 1 ,Numan AKYOL 1 ,Tahir KARAŞAHİN 1 ,Muharrem SATILMIŞ 1

1 Lalahan Hayvancılık Merkez Arastırma Enstitüsü – ANKARA - The aim of the study was to investigate the effect of vitrification on in vitro produced bovine embryos. For this purpose, oocytes were provided from ovaries of slaughtered cows and 7 days after start of embryo culture 100 good quality in vitro produced bovine embryos were frozen by vitrification method. Vitrification solutions were composed as VS1 (0.1 M Sucrose + 0.1 M Xylose + 1% Poly Ethylene Glycol + 10% Glycerol), VS2 (0.2 M Sucrose + 0.2 M Xylose + 2% Poly Ethylene Glycol +10 % Glycerol + 10% Ethylene Glycol) and VS3 (0.3 M Sucrose + 0.3 M Xylose + 3% Poly Ethylene Glycol + 20% Glycerol + 20% Ethylene Glycol). The processed embryos with vitrification solutions were frozen in liquid nitrogen after placed into straws. The straws were thawed by holding in air for 5-6 sec and in 20ºC water. Embryos were transferred 0.5 M and 0.25 M sucrose solutions for devitrification procedure respectively. Then, the embryos were transferred into D-PBS (with 20% calf serum) for morfological evaluation. TCM-199 + 0.1mM Β-Mercaptoethanol + 20% Foetal Calf Serum (FCS) composition was used as culture media. For in vitro survival control, embryos were incubated in the conditions with regard to 5% CO2, at 38.5ºC and 99% humidity for 24-48 hours. 85 intacted embryos were obtained from 100 embryos after thawing in Phospate Buffer Solution. The re-expansion and hatching rate of blastocysts were 35%. So, in vitro produced and vitrified bovine embryos survived with rate of 35%. As a result, in vitro produced embryos were frozen easily by vitrification method and provided satisfactory level for survival. Keywords : Embryo, bovine, vitrification, cryoprotectant