Temporal sensitivity of bovine embryos to culture environment after fertilization and the implications for blastocyst quality.
The aim of this study was to examine the temporal sensitivity of bovine embryos to culture environment after fertilization to determine which period, if any, is most critical in determining blastocyst quality. Bovine zygotes produced in vitro were divided into six groups and cultured either in vitro (in synthetic oviductal fluid, SOF), in vivo (in the ewe oviduct) or in a combination of both systems. Development to the blastocyst stage, the ability of the blastocysts to withstand cryopreservation and the relative abundance of several gene transcripts were examined. Culture in SOF for either 2 or 4 days, followed by subsequent culture in the ewe oviduct, resulted in a significantly lower yield of blastocysts than did all other methods, the effect being most marked in embryos that were cultured in SOF for 4 days. In contrast, culture in vivo for the first 2 or 4 days after fertilization followed by culture in vitro did not have such a marked effect on blastocyst development. Blastocysts produced after culture in the oviduct for 6 days had the highest rates of survival over 72 h after warming (100% survival at 24 h; >95% survival at 72 h). The embryos that spent the last 4 days of culture in vivo also had relatively high rates of survival (100% at 24 h, 73.7% at 72 h). Blastocysts produced entirely in SOF had very low rates of survival after vitrification, with <40% viable at 24 h and <20% survival at 72 h. Blastocysts derived from embryos that spent the first 2 days in vivo and the last 4 days in vitro had the lowest rates of survival (6.7%), whereas those that spent the last 2 days only in SOF had intermediate rates of survival (40.6%). These differences were reflected in the relative abundance of transcripts for the Bax gene.
Lonergan
P
P
Department of Animal Science and Production and Conway Institute for Biomedical and Biomolecular Research, University College Dublin, Lyons Research Farm, Newcastle, County Dublin, Ireland. pat.lonergan@ucd.ie
Rizos
D
D
Kanka
J
J
Nemcova
L
L
Mbaye
A M
AM
Kingston
M
M
Wade
M
M
Duffy
P
P
Boland
M P
MP
D000818:Animals; D001755:Blastocyst / Q000502:physiology*; D001826:Body Fluids / Q000378:metabolism; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods; D015925:Cryopreservation / Q000379:methods; D017077:Culture Media, Conditioned; D004624:Embryo Transfer; D005314:Embryonic and Fetal Development; D005187:Fallopian Tubes / Q000378:metabolism; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods; D005313:Fetal Death; D015870:Gene Expression; D011518:Proto-Oncogene Proteins / Q000235:genetics; D019253:Proto-Oncogene Proteins c-bcl-2; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D015053:Zygote / Q000166:cytology* / Q000378:metabolism; D051028:bcl-2-Associated X Protein
D017077:Culture Media, Conditioned; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein