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1. Reprod Fertil Dev. 2011 Dec;24(1):160.

95 sheep zygotes cultured with hyaluronan, bovine serum albumin, or serum and
their effects on prenatal and postnatal embryo production and development.

Ali AB.

Department of Animal Biology, University of Sassari, Sassari, Italy.

Mammalian pre-implantation embryos are sensitive to their environment and
numerous publications suggest that post-fertilization culture is the most
critical period affecting embryo quality and subsequent embryo development. This
study was conducted to examine the viability, twining, sex ratio and birthweight
(BW) after transfer of fresh and vitrified in vitro-produced sheep embryos
derived from oocytes cultured in a semi- or defined medium supplemented with
fatty-acid-free bovine serum albumin (BSA) with or without hyaluronan (HA) versus
fetal bovine serum (FBS) as a control. A total of 649 oocytes were matured in
TCM-199 containing 2mM glutamine, 4mg mL(-1) BSA, 100μM cysteamine, 0.3mM sodium
pyruvate, 1μg mL(-1) oestradiol-17β and fertilized with fresh semen prepared with
SOF, enriched with 20% oestrus sheep serum. Presumptive zygotes were cultured for
48h in SOF supplemented with 1% BME-essential and 1% MEM-nonessential amino
acids, 1mM glutamine and 8mg mL(-1) BSA. On the 3rd day of culture, presumptive
zygotes were divided randomly into 3 groups: (1) BSA+HA, medium supplemented with
8mg mL(-1) BSA and 6mg mL(-1) HA, (2) BSA, medium supplemented with 8mg mL(-1)
BSA only, or (3) control, medium supplemented with 5% charcoal-stripped fetal
bovine serum (FBS). On the 5th day of culture, media were replaced accordingly in
all groups. Embryos that reached the expanded blastocyst stage by 6 or 7
days/group were recorded and transferred as fresh or vitrified. The cleavage
rate, blastocyst formation, pregnancies (PR) at 40, 60 and 80 days, lambing,
twinning, sex ratio and BW were all recorded. Among all groups, no significant
differences were observed in cleavage and blastocyst rates. The proportion of
zygotes that developed to blastocysts at 6 days was significantly higher after
culture with BSA+HA vs BSA and control (67/92, 72.8%), (46/118, 39.0%) and
(29/72, 40.3%), respectively. Compared with the control group, vitrified BSA
blastocysts were associated with increased (P<0.05) PRs after 60 and 80 days and
lambing rates. In comparison, there were no significant differences in pregnancy
and lambing rate after transfer of fresh blastocysts. There was a positive
correlation between age of embryo at transfer and higher BW. The mean BWs were
higher after transfer of 7-day vs 6-day embryos (3.95 and 3.37kg), respectively.
However, there were no significant differences in twining and sex ratio after
transfer fresh/or vitrified treated embryos versus control group. In conclusion,
the results indicate that sheep zygotes cultured with BSA+HA had an increased
development to blastocysts at 6 days, but the rate of blastocyst formation was
not increased, PR and lambing rate were comparable with the BSA and control
groups. In addition, transfer of vitrified embryos produced in the presence of
BSA significantly increased the PR at 60, 80 days and lambing rate versus control

PMID: 22394817 [PubMed - in process]