Author/Authors :
Hosseini، Sayyed Morteza نويسنده Clinical and Experimental Embryology Department, Cell Sciences Research Centre, Royan Institute (Isfahan Campus), ACECR, Tehran, Iran , , Moulavi، Fariba نويسنده , , Hajian، Mahdi نويسنده , , Abedi، Parvaneh نويسنده , , Forouzanfar، Mohsen نويسنده , , Ostad Hosseini، Somayyeh نويسنده , , Hosseini، Laleh نويسنده , , Pirestani، Akbar نويسنده , , Ghasemzadeh Nava، Hamid نويسنده Clinical Science Department, Faculty of Veterinary Medicine , University of Tehran, Tehran, Iran , , Tajik، Parviz نويسنده , , Shahverdi، abdol Hossein نويسنده , , NASR ESFAHANI، MOHAMMAD HOSSEIN نويسنده ,
Abstract :
Background: The aim of this study was to establish a cell-free sequential culture system that can
support high levels of in vitro embryo development and blastocyst formation from bovine zygotes.
To this end, this investigation was carried out to evaluate the effects of glucose, serum and EDTA
on bovine zygote in vitro development.
Materials and Methods: Bovine presumptive zygotes were derived from oocytes matured, and
fertilized in vitro and cultured in synthetic oviductal fluid sequential medium in a two-steps manner;
SOF 1 for the first 3 days and SOF 2 for the second 5-6 days of in vitro embryo development. In
order to evaluate the effect of different modifications of the basic medium on embryo development,
glucose was added to the second phase (SOF A), serum was added to the first phase (SOF C) and
EDTA alone (SOF D) or in combination with serum (SOF E) was added into the first phase of in
vitro embryo culture. The results of each composition were compared with each other and with the
results of embryo development in TCM199 vero cell co-culture system.
Results: Glucose addition to the second phase of embryo culture, improved the developmental
competency; however, the differences were not significant. Serum addition to the first phase of
embryo culture, significantly improved the developmental competency of embryos beyond the
cleavage stage, compared to all the treatment and TCM199 co-culture groups. EDTA supplementation
of culture medium, either alone or in combination with serum, significantly inhibits the embryo
development beyond the morula stage.
Conclusion: The results indicated that culture of bovine presumptive zygotes in two steps cell-free
culture system, can support embryo development, and addition of serum throughout the culture and
glucose to the second step significantly increased overall developmental competency compared to
TCM199 co-culture system.
