Effect of the post-fertilization culture environment on the incidence of chromosome aberrations in bovine blastocysts.
We have previously shown that the postfertilization embryo culture environment has a significant influence on the quality of the resulting bovine blastocyst measured in terms of its cryotolerance and relative abundance for several developmentally important gene transcripts. Using three different culture conditions known to produce blastocysts of differing quality, the objective of this study was to examine whether the postfertilization culture environment had an effect on the incidence of mixoploidy in bovine blastocysts. Presumptive zygotes, produced by in vitro maturation and fertilization, were cultured in vitro in synthetic oviduct fluid (SOF) medium in the absence or presence of fetal calf serum (FCS), or in vivo in the ewe oviduct. Blastocysts were recovered from the three systems at Day 7 and the incidence of mixoploidy was assessed using fluorescence in situ hybridization with chromosome 6- and chromosome 7-specific probes. A total of 10 025 nuclei were scored in 122 blastocysts. The frequency of normal, diploid, blastocysts was 8.8%, 21.4%, and 34.8% in embryos derived from culture in SOF+FCS, SOF, and the ewe oviduct, respectively, the remainder showing some degree of mixoploidy. The incidence of mixoploidy was apparently not related to the presence of serum; omission of serum from SOF resulted in a reduction in the incidence of mixoploidy (91.2% vs. 78.6%), although this difference was not significant. Culture in vivo, however, resulted in a significant (P < 0.01) reduction in the incidence of mixoploidy compared with culture in vitro in the presence of serum (65.2% vs. 91.2%, respectively). Among the mixoploid blastocysts, the majority contained less than 10% polyploid cells, irrespective of culture group (SOF, 69.7%; SOF+FCS, 64.5%; ewe oviduct, 60.0%). More than one type of polyploidy was frequently observed in mixoploid blastocysts. Overall, diploidy-triploidy was the most frequent abnormality, but diploid-tetraploid and diploid-triploid-tetraploid mosaics were also observed. A significantly higher proportion (P < 0.05) of blastocysts derived from SOF+FCS had more than one type of abnormality (80.6%, 25/ 31) compared with those derived from SOF (45.4%, 15/33) or in vivo culture (53.3%, 16/30). In conclusion, the postfertilization culture environment of the developing embryo can affect the incidence and severity of mixoploidy in the resulting blastocyst.
Lonergan
Patrick
P
Department of Animal Science and Production, University College Dublin, Lyons Research Farm, Newcastle, County Dublin, Ireland. pat.lonergan@ucd.ie
Pedersen
Hanne G
HG
Rizos
Dimitrios
D
Greve
Torben
T
Thomsen
Preben D
PD
Fair
Trudee
T
Evans
Alex
A
Boland
Maurice P
MP
D000818:Animals; D001755:Blastocyst / Q000166:cytology*; D002417:Cattle / Q000196:embryology*; D002467:Cell Nucleus; D002869:Chromosome Aberrations / Q000139:chemically induced / Q000662:veterinary*; D033481:Chromosomes, Mammalian / Q000145:classification; D003470:Culture Media / Q000633:toxicity; D046149:Embryo Culture Techniques / Q000379:methods*; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D009865:Oocytes / Q000166:cytology; D011003:Ploidies; D011897:Random Allocation; D012756:Sheep
D003470:Culture Media