Cryopreservation of Bos taurus vs Bos indicus embryos: are they really different?
Cryopreservation with storage at very low temperatures is essential to make full use of this technology for both biological and commercial reasons. However, most mammalian cells will die if exposed to these temperatures unless they are exposed to cryoprotectant solutions and cooled and warmed at specific rates. Lowering temperature below 0 degree C introduces the risk of intracellular ice formation, which likely increases rapidly as the temperature falls. Evidence indicates that ice formation during cooling can cause significantly more damage than ice formation during warming. Comparisons of the toxicity of various cryoprotectants indicated that ethylene glycol (EG) is a nontoxic compound for murine and bovine embryos. The 3.6 M EG solution resulted in similar high survival rates when compared with nonfrozen embryos; deleterious effects of high concentrations of EG became apparent at 7.2 M. The use of EG provides a nontoxic method for the rapid and simplified controlled freezing of in vivo bovine compact morulae-early blastocyst, avoiding the risk of injury caused by high concentrations of cryoprotectants usually required for vitrification. However, in vivo embryos used for freezing and thawing require further studies to understand the ultrastructural changes during the freezing procedure with EG as the single cryoprotectant, especially between Holstein and Nelore cows. This paper describes the ultrastructure of bovine compact morulae-early blastocysts derived by in vivo methods from Holstein and Nelore cows to investigate the fresh morphology as well as that after exposure to cryoprotectant and cryopreservation by conventional slow freezing, quick freezing (nitrogen vapor), and vitrification.
Visintin
J A
JA
Department of Animal Reproduction, Faculty of Veterinary Medicine, Institute of Biomedical Sciences, University of Sao Paulo, Sao Paulo, 05508-000, Brazil.
Martins
J F P
JF
Bevilacqua
E M
EM
Mello
M R B
MR
Nicácio
A C
AC
Assumpção
M E O A
ME
D000818:Animals; D001755:Blastocyst / Q000187:drug effects / Q000648:ultrastructure*; D002417:Cattle / Q000145:classification / Q000196:embryology* / Q000502:physiology*; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000633:toxicity*; D004622:Embryo, Mammalian / Q000166:cytology / Q000187:drug effects / Q000648:ultrastructure; D005260:Female; D005615:Freezing; D008297:Male; D008854:Microscopy, Electron / Q000662:veterinary; D009028:Morula / Q000187:drug effects / Q000648:ultrastructure*; D013997:Time Factors
D003451:Cryoprotective Agents