Abstract
Evaluation of quality and viability of bovine and rabbit preimplantation embryos following cryopreservation was the objective of this study. Bovine embryos of Holstein breed (n= 88) at the morula-early blastocyst stage (on the 7 th day after the first insemination) and rabbit embryos (n= 135) from New Zealand breed at the morula stage (90–92 hpc) were cryopreserved by the two-step vitrification procedure. Following thawing the embryos of both cattle and rabbit were cultured for 48 hours in order to reach expanded blastocyst stage, and then were analyzed for the developmental rate and viability (embryo cell number and incidence of the dead cells). The results demonstrate that cryopreservation can only slightly affect embryo viability and quality do not compromising their further development. Therefore, vitrification techniques tested in our study can be used for cryopreservation of embryos of national cattle and rabbit breeds for the purpose of long-term storage of embryos in the animal gene bank.
Highlights
Cryopreservation of preimplantation embryos is an important tool for maintaining animal genetic sources by increasing the usage of reproductive potential of genetically valuable animals
Following thawing the embryos of both cattle and rabbit were cultured for 48 hours in order to reach expanded blastocyst stage, afterwards these embryos were analyzed for the developmental rate, embryo cell number and incidence of the dead cells
Both fresh and frozen-thawed embryos were cultured since the morula stage up to 48 hours, when it was expected that most of embryos can reach either blastocyst or expanded blastocyst stage
Summary
Cryopreservation of preimplantation embryos is an important tool for maintaining animal genetic sources by increasing the usage of reproductive potential of genetically valuable animals. Cryopreservation procedures may negatively affect survival of gametes and embryos as a consequence of damages to the cell and even cell death (apoptosis or necrosis). Determination of these influences may have a great importance for the explanation of failures in reproductive processes. As a functional criteria for the evaluation of embryo viability after cryopreservation, a post-thaw cleavage up to blastocyst stage (Popelkova et al, 2005; Makarevich et al 2008), embryo cell number and embryo diameter (Popelkova et al, 2009), proliferation (PCNA) index (Markkula et al, 2001) or number of apoptotic (TUNEL) cells (Marquez-Alvarado et al, 2004; Makarevich et al, 2008) and the state of actin cytoskeleton (Tharasanit et al 2005; Makarevich et al, 2008) have been used
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