| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
We have previously shown that while the intrinsic quality
of the oocyte is the main factor affecting blastocyst
yield during bovine embryo development in vitro, the main
factor affecting the quality of the blastocyst is the
post-fertilization culture conditions. Therefore, any
improvement in the quality of blastocysts produced in
vitro is likely to derive from the modification of the
post fertilization culture conditions. The objective of
this study was to examine the effect of the presence or
absence of serum and the concentration of bovine serum
albumin (BSA) during the period of embryo culture in vitro
on 1) cleavage rate, 2) the kinetics of embryo
development, 3) blastocyst yield, and 4) blastocyst
quality, as assessed by cryotolerance and gene expression
patterns. The quantification of all gene transcripts was
carried out by real time quantitative RT-PCR. Bovine
blastocysts from four sources were used: 1) in vitro
culture in synthetic oviduct fluid (SOF) supplemented with
3 mg/ml BSA and 10% fetal calf serum (FCS), 2) in vitro
culture in SOF + 3 mg/ml BSA in the absence of serum, 3)
in vitro culture in SOF + 16 mg/ml BSA in the absence of
serum, and 4) in vivo blastocysts.
There was no difference in overall blastocyst yield at Day
9 between the groups. However, significantly more
blastocysts were present by Day 6 in the presence of 10 %
serum (20.0%) compared with 2 mg/ml BSA (4.6%, P<0.001) or
16 mg/ml BSA (11.6%, P<0.01). By Day 7, however, this
difference had disappeared. Following vitrification, there
was no difference in survival between blastocysts produced
in the presence of 16 mg/ml BSA or those produced in the
presence of 10% FCS; the survival of both groups was
significantly lower than the in vivo controls at all time
points and in terms of hatching rate. In contrast,
survival of blastocysts produced in SOF + 3mg/ml BSA in
the absence of serum was intermediate with no difference
remaining at 72 h when compared to in vivo embryos.
Differences in relative mRNA abundance amongst the two
groups of blastocysts analysed were found for genes
related to apoptosis (Bax), oxidative stress (MnSOD,
CuZnSOD and SOX), communication through gap junctions
(Cx31 and Cx43), maternal recognition of pregnancy
(IFN-
) and differentiation and implantation (LIF and
LR-ß). The presence of serum during the culture
period resulted in a significant increase in the level of
expression of MnSOD, SOX, Bax, LIF and LR-ß. The
level of expression of Cx31 and Cu/ZnSOD was also
increased although the difference was not significant. In
contrast, the level of expression of Cx43 and IFN-
was decreased in the presence of serum.
In conclusion, using a combination of measures of
developmental competence (cleavage and blastocyst rates)
and qualitative measures such as cryotolerance and
relative mRNA abundance to give a more complete picture of
the consequences of modifying medium composition on the
embryo we have shown that conditions of post fertilization
culture, in particular, the presence of serum in the
medium, can affect the speed of embryo development and the
quality of the resulting blastocysts. The reduced
cryotolerance of blastocysts generated in the presence of
serum is reflected in deviations in the relative abundance
of developmentally important gene transcripts. Omission of
serum during the post fertilization culture period can
significantly improve the cryotolerance of the blastocysts
to a level intermediate between serum-generated
blastocysts and those derived in vivo. The challenge now
is to try and bridge this gap.
This article has been cited by other articles:
|
|
 |
|
  S L Rodriguez-Zas, K Schellander, and H A Lewin Biological interpretations of transcriptomic profiles in mammalian oocytes and embryos Reproduction, February 1, 2008; 135(2): 129 - 139. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. R. Herrick, J. B. Bond, G. M. Magarey, H. L. Bateman, R. L. Krisher, S. A. Dunford, and W. F. Swanson Toward a Feline-Optimized Culture Medium: Impact of Ions, Carbohydrates, Essential Amino Acids, Vitamins, and Serum on Development and Metabolism of In Vitro Fertilization-Derived Feline Embryos Relative to Embryos Grown In Vivo Biol Reprod, May 1, 2007; 76(5): 858 - 870. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. J. Stokes, J. A. Hawkhead, R. K. Fawthrop, H. M. Picton, V. Sharma, H. J. Leese, and F. D. Houghton Metabolism of human embryos following cryopreservation: Implications for the safety and selection of embryos for transfer in clinical IVF Hum. Reprod., March 1, 2007; 22(3): 829 - 835. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. El-Sayed, M. Hoelker, F. Rings, D. Salilew, D. Jennen, E. Tholen, M.-A. Sirard, K. Schellander, and D. Tesfaye Large-scale transcriptional analysis of bovine embryo biopsies in relation to pregnancy success after transfer to recipients Physiol Genomics, December 13, 2006; 28(1): 84 - 96. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Nganvongpanit, H. Muller, F. Rings, M. Hoelker, D. Jennen, E. Tholen, V. Havlicek, U. Besenfelder, K. Schellander, and D. Tesfaye Selective degradation of maternal and embryonic transcripts in in vitro produced bovine oocytes and embryos using sequence specific double-stranded RNA. Reproduction, May 1, 2006; 131(5): 861 - 874. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Sagirkaya, M. Misirlioglu, A. Kaya, N. L First, J. J Parrish, and E. Memili Developmental and molecular correlates of bovine preimplantation embryos. Reproduction, May 1, 2006; 131(5): 895 - 904. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D Corcoran, T Fair, S Park, D Rizos, O V Patel, G W Smith, P M Coussens, J J Ireland, M P Boland, A C O Evans, et al. Suppressed expression of genes involved in transcription and translation in in vitro compared with in vivo cultured bovine embryos. Reproduction, April 1, 2006; 131(4): 651 - 660. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Lonergan, H. G. Pedersen, D. Rizos, T. Greve, P. D. Thomsen, T. Fair, A. Evans, and M. P. Boland Effect of the Post-Fertilization Culture Environment on the Incidence of Chromosome Aberrations in Bovine Blastocysts Biol Reprod, October 1, 2004; 71(4): 1096 - 1100. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Fernandez-Gonzalez, P. Moreira, A. Bilbao, A. Jimenez, M. Perez-Crespo, M. A. Ramirez, F. R. De Fonseca, B. Pintado, and A. Gutierrez-Adan Long-term effect of in vitro culture of mouse embryos with serum on mRNA expression of imprinting genes, development, and behavior PNAS, April 20, 2004; 101(16): 5880 - 5885. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Fair, A. Gutierrez-Adan, M. Murphy, D. Rizos, F. Martin, M. P. Boland, and P. Lonergan Search for the Bovine Homolog of the Murine Ped Gene and Characterization of Its Messenger RNA Expression During Bovine Preimplantation Development Biol Reprod, February 1, 2004; 70(2): 488 - 494. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X.-S. Cui, Y.-J. Jeong, H.-Y. Lee, S.-H. Cheon, and N.-H. Kim Fetal bovine serum influences apoptosis and apoptosis-related gene expression in porcine parthenotes developing in vitro Reproduction, January 1, 2004; 127(1): 125 - 130. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. E. Farin, P. W. Farin, and J. A. Piedrahita Development of fetuses from in vitro-produced and cloned bovine embryos J Anim Sci, January 1, 2004; 82(13_suppl): E53 - 62. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Lonergan, D. Rizos, A. Gutierrez-Adan, P.M. Moreira, B. Pintado, J. de la Fuente, and M.P. Boland Temporal Divergence in the Pattern of Messenger RNA Expression in Bovine Embryos Cultured from the Zygote to Blastocyst Stage In Vitro or In Vivo Biol Reprod, October 1, 2003; 69(4): 1424 - 1431. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
