Likewise, Vajta et al. [37] demonstrated severe degenerative changes in cells of in vitro produced bovine embryos immediately after warming. But during the subsequent 4 h culture evident signs of regeneration were observed, and after 24 h only slight signs of injury could still be seen. In preantral follicle oocytes, vitrification significantly affected mitochondrial inner membrane potential [10], but mitochondrial activity was recovered after 12 days in culture. Similarly, human blastocysts had their respiratory rate lowered or

even absent after vitrification/warming, only detected again after 24 h [40]. Undoubtedly, one hour of IVC was not enough to allow metabolic recovery in the present study. How long would it take to mitochondrial activity to be restored in these cryopreserved embryos remains a question. Mitochondrial malfunction may be caused by decline in the mitochondrial Alpelisib research buy membrane

potential and disruption of mitochondrial membrane. While the first is often reversible [10], [29] and [40], membrane disruption is a Y-27632 price more critical damage. Comparing mitochondrial ultrastructure of fresh and cryopreserved embryos, swollen mitochondria were more frequent in cryopreserved embryos. However, most mitochondria from embryos grade I and II post-cryopreservation presented typical ultrastructure. No rupture of mitochondrial membranes was seen on grade I and II embryos in

this study. Higher degrees of mitochondrial swelling were observed in previous studies on cryopreserved grade I and II sheep embryos [2] and [5]. Mitochondrial swelling is also commonly described in cryopreserved oocytes [14], [16] and [23]. Using in vitro produced embryos and similar procedures of slow freezing and vitrification Bettencourt et al. [3] achieved satisfactory pregnancy rates of 68.4% and 54.6% on day 45, respectively. This shows that some Temsirolimus solubility dmso ultrastructural changes observed on transferable embryos after cryopreservation are reversible, and embryos can fully recover. Besides playing a role in organelle organization the primary function of actin filaments is acting on intercellular junctions during the compaction process and to maintain structural integrity during the initial embryo stages [18]. The layout of actin filaments during the transition stage from morulae to initial blastocyst is justified by asymmetric division, polarization and differentiation of ICM and trophoblastic cells [27]. Cryopreserved embryos were characterized by mild to severe disorganization of actin filaments. Better quality embryos (grade I and II) presented small cytoskeleton damage. Cryopreserved grade III embryos showed a high level of cytoskeleton disorganization, independent of the cryopreservation treatment.