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| Status |
Public on Mar 23, 2017 |
| Title |
Differences prevailing in bovine in vitro produced blastocysts in the view of their post-transfer phenotypes three days ahead of implantation |
| Organism |
Bos taurus |
| Experiment type |
Expression profiling by array
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| Summary |
Embryo transfer is largely used in cattle and classically performed at D7 (or D8) using unsorted D7 (or D8) blastocysts produced in vivo, or in vitro in defined media without serum or feeders. Outdated systems including serum and co-culture were however of interest for research purposes. We thus wondered whether embryos that would form a blastocoel at different times after fertilisation (D6 to D8) and stay in culture for up to 2 additional days (D6+1, D6+2, D7+1) would equally develop in vivo after temporary transfer to oestrus-synchronised recipients. Globally alike, those that survived up to D18 reached primitive streak stages and elongated to filamentous sizes similarly to in vivo (D18) or in vitro controls (classical D7-T7). Recovery rate differed between D6 and D8 embryos that were immediately transferred (58 vs 25% in D6-T6 vs D8-T8). With a reduced but intermediate survival (33%), the D6 embryos that stayed 2 more days in culture produced 7 times more IFN-tau at D18 than the immediately transferred D6 embryos. At the end of the culture, D6+2 embryos also displayed the higher number of differences with the D6 blastocysts. A “+1” phenotype emerged from the D6+1 and D7+1 embryos, that shared a larger gene set enrichment than the blastocysts they derived from, possible sign of a similar adaption to the in vitro environment. To the best of our knowledge, this is the first time that this culture system (B2, serum, co-culture) is used to study its impacts on the embryonic transcriptome prior to transfer. Initially reputed as beneficial to produce more expanding and hatching blastocysts, this culture system generated blastocysts that all dissembled in vivo developed ones (D7). Despite a loss of 40 to 60% in the two weeks after ET, no dying (vs surviving) signature was detectable in any of the transferred groups (D6, D6+2, D7+1, D8); was it rather a matter of developmental “pause”? Whether molecular differences prevailing to transfer partly reflected those induced by unfavourable conditions (or diapause) was therefore assessed.
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| Overall design |
After in vitro fertilization, bovine zygotes were cultured in batch of 25 embryos in 50 µl drops, at 39°C with 5% O2, 5% CO2 and 90% N2 until blastocoel formed at day 6 (condition “6”) or day 8 (condition “8”). One and two days after day 6, early formed blastocysts were also collected, not as “day 8 forming ones” but as “cultured 1 or 2 more days” (condition “6+1 and 6+2”). One days after day 7, early formed blastocysts were also collected, not as “day 8 forming ones” but as “cultured 1 more day” (condition “7+1”)
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| Contributor(s) |
Hue I, Dufort I, Vitorino Carvalho A, Laloe D, Degrelle S, Viebahn C, Sirard M |
| Citation(s) |
30444718 |
| Submission date |
Mar 22, 2017 |
| Last update date |
Nov 27, 2018 |
| Contact name |
Marc-André Sirard |
| E-mail(s) |
Marc-Andre.Sirard@fsaa.ulaval.ca
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| Organization name |
Université Laval
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| Department |
Sciences Animales
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| Street address |
Offfice 2732, 2440 Hochelaga Blvd.
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| City |
Québec City |
| State/province |
Quebec |
| ZIP/Postal code |
G1V 0A6 |
| Country |
Canada |
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| Platforms (1) |
| GPL13226 |
Agilent-028298 Embryogene Bovine 45K microarray |
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| Samples (18)
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| Relations |
| BioProject |
PRJNA380117 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE96925_RAW.tar |
21.2 Mb |
(http)(custom) |
TAR (of TXT) |
| Processed data included within Sample table |
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