85 Elongation of trophoblastic vesicles between Days 15 and 18 in cattle

Abstract

Once formed, bovine blastocysts differentiate while growing exponentially from 150-200 μm (Days 7 or 8) to 200-250 mm (Days 17 to 18; Sandra et al. 2017 Annu. Rev. Anim. Biosci. 5, 205-228). Thus, the length of the conceptus doubles every day between Days 9 and 16 (Berg et al. 2010 Theriogenology 73, 250-260); however, this was observed on whole conceptuses. The objective of the current study was to test whether this elongation rate is similar when the embryonic disc has been excised. Six heifers were used to produce Day-15 conceptuses, either fully developed in vivo or developed in vivo for a week after embryo transfer of Day 8 in vitro-produced blastocysts. Day 15 conceptuses were recovered, measured, and cut into pieces to produce trophoblastic vesicles (TV; Heyman et al. 1984 J. Reprod. Fertil. 70, 533-540) of 4 ± 0.07 or 4.4 ± 0.65 mm long (mean ± standard error of the means) for in vivo- or in vitro-produced TV, respectively. All TV were transferred into oestrus-synchronized recipients (5 heifers and 1 cow). Each female received 8-9 TV so that a total of 24 in vivo-derived and 26 in vitro-produced TV were transferred in utero for a period of 3 days. The TV originating from different Day-15 conceptuses were mixed at the time of transfer, so that each recipient received the TV from different origins (conceptus and donor cow). Transcervical collection was used at Day 15 and 18 for conceptus and TV recovery (Richard et al. 2015 Theriogenology 83, 1101-1109). At Day 18, TV elongation size was analysed (mean ± standard error of the means) by unpaired t-test using GraphPad Prism software (GraphPad Inc., San Diego, CA, USA). At Day 15, conceptuses from the in vivo and in vitro groups displayed different sizes and length variabilities (24-32 v. 2-24 mm, respectively). At Day 18, TV recovery rate was 79% for the in vivo- v. 62% for the in vitro-derived group and mean elongation rate (over 3 days) was ×5.4 (minimum = 2.5, maximum = 10) v. ×7.6 (minimum = 0.7, maximum = 20.5), respectively. There was no significant difference for TV size between groups at Day 18 (21.75 ± 2.24 mm v. 33.38 ± 11.63 mm, respectively). Altogether, the variability in length at Day 15 was previously reported, the difference in TV recovery between in vivo and in vitro groups reached 17% and was similar to the loss of 11% that occurs in the first week after classical embryo transfer. In opposition to studies where in vitro-produced conceptuses were shorter than in vivo-developed ones, in vivo and in vitro groups of TV likely followed similar growth. Whether this reflects a normal growth awaits further studies.