title: Developmental kinetics of the first cell cycles of bovine in vitro produced embryos in relation to their in vitro viability and sex.
abstract: The development of bovine IVP-embryos was observed in a time-lapse culture system to determine cell cycle lengths of 1) embryos that developed into compact morulae (CM) or blastocysts (BL) within 174 h after insemination (viable), 2) embryos that arrested during earlier stages (nonviable) and 3) male and female embryos. In 4 replicates, inseminated oocytes were cultured on a microscope stage in 3 to 4 groups on a granulosa cell monolayer in supplemented TCM 199. Images were sequentially recorded and stored at 30-min intervals. All embryos that could be identified throughout the culture period were included (n = 392), and the times of cleavage events noted. After culture, 100 CM or BL were randomly selected for sexing by PCR. BL developed equally well in the time-lapse and control culture systems (36 vs 38%). The respective lengths of the first 4 cell cycles of viable embryos were 32.0 +/- 3.9, 8.8 +/- 1.6, 10.8 +/- 4.7 and 47.7 +/- 11.8 h. The subsequent intervals between the 9- to 16-cell, early morula, CM and BL stages lasted 16.2 to 18.2 h. Blastomeres of 2-, 4- and 8-cell embryos cleaved asynchronously with < 1, 2.6 +/- 2.5 and 9.2 +/- 4.5 h intervals, respectively, between the first and last blastomere to cleave. The interval from insemination to tight compaction and formation of a blastocoel was 128.4 +/- 10.7 and 145.8 +/- 12.5 h, respectively. The first 3 cell cycles were approximately 3 h shorter (P < 0.1) while the fourth cycle was 5 h shorter (P = 0.06) for the viable vs nonviable embryos. On this basis it was possible to define time windows in which the proportion of viable 2-, 3- to 4-, 5- to 8- and 9- to 16- cell embryos were at their highest. No differences were found between the cleavage intervals of male and female embryos. We conclude 1) that the time-lapse culture system allows for detailed observation of the developmental kinetics of several embryo groups at the same time, and 2) that these embryos can be manipulated at the end of culture, thus allowing a linkage between early cleavage events and other developmental parameters such as embryo sex or viability after transfer.
authors.affiliation: Embryo Technology Center, Danish Institute of Agricultural Sciences, Tjele, Denmark.
authors.forename: P
authors.initials: P
authors.lastname: Holm
authors.forename: N N
authors.initials: NN
authors.lastname: Shukri
authors.forename: G
authors.initials: G
authors.lastname: Vajta
authors.forename: P
authors.initials: P
authors.lastname: Booth
authors.forename: C
authors.initials: C
authors.lastname: Bendixen
authors.forename: H
authors.initials: H
authors.lastname: Callesen
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000502:physiology; D002417:Cattle / Q000196:embryology*; D002970:Cleavage Stage, Ovum; D046508:Culture Techniques; D004624:Embryo Transfer; D005314:Embryonic and Fetal Development; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D007700:Kinetics; D008297:Male; D009028:Morula / Q000502:physiology; D012727:Sex Characteristics; D013997:Time Factors