Vitrification of in vitro cultured porcine two-to-four cell embryos
Vitrification of in vitro cultured porcine two-to-four cell embryos
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DOI：
10.1016/j.theriogenology.2007.05.039
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发表时间：
2007-07-15
期刊：
影响因子：
2.8
通讯作者：
Martinez, E. A.
中科院分区：
文献类型：
--
作者：
Cuello, C.;Gil, M. A.;Martinez, E. A.
本实验的目的是评估2 - 4细胞猪胚胎到囊胚期体外培养5天对其玻璃化和加热存活能力的影响。为了提高冷却速度，采用超细开拉吸管和vitm - master (R)技术进行玻璃化。在第2天（DO =发情开始）从断奶母猪（n = 11）收集2 - 4个细胞胚胎。部分胚胎（N = 63）采集后3 d内玻璃化，加热培养120 h （V2组）。此外，81个2 - 4细胞胚胎培养96个，以获得囊胚；然后玻璃化，加热，培养24 h （VB组；N = 65）。剩余的2 - 4个细胞胚胎作为对照，因此没有玻璃化（对照胚胎；N = 70），但在体外培养120小时。在培养过程中评估V2， VB和对照胚胎的发育进展和形态。所有胚胎（V2， VB和对照）在发育当天固定，以评估卵裂球总数。V2胚的成活率和囊胚形成率非常低（分别为9.6 +/- 0.7%和3.2 +/- 0.5%）。VB胚的成活率和孵化率（75.0 +/- 0.69%和33.6 +/- 0.13%）低于对照胚（89.1 +/- 0.8%和47.5 +/- 0.12%）（p < 0.001）。孵化后的VB胚总细胞数（70.3 +/- 4.5）低于孵化后的对照胚（90.6 +/- 3.2）（p < 0.01）。扩大后的VB囊胚与对照囊胚无明显差异。总之，2 - 4细胞猪胚胎体外培养的囊胚可以使用SOPS吸管和vitm - master (R)成功地玻璃化。(c) 2007 Elsevier Inc.。版权所有。
The objective of this experiment was to evaluate the effect of a 5-day period of in vitro culture of two-to-four cell porcine embryos up to the blastocyst stage on their ability to survive vitrification and warming. In order to increase the cooling rate, superfine open pulled straws and Vit-Master (R) technology were used for vitrification. Two-to-four cell embryos were collected from weaned sows (n = 11) on day 2 (DO = onset of estrus). Some embryos (N = 63) were vitrified within 3 It after collection, warmed and cultured for 120 h (Group V2). Additionally, 81 two-to-four cell embryos were cultured for 96 It in order to obtain blastocysts; these were then vitrified, warmed and cultured for 24 h (Group VB; N = 65). The remaining two-to-four cell embryos were used as controls and thus not vitrified (control embryos; N = 70) but were cultured in vitro for 120 h. The V2, VB and control embryos were evaluated for their developmental progression and morphology during culture. All embryos (V2, VB and controls) were fixed on the same day of development in order to assess the total number of blastomeres. The survival and blastocyst formation rates obtained from V2 embryos were very poor (9.6 +/- 0.7% and 3.2 +/- 0.5%, respectively). The survival and hatching rates of VB embryos (75.0 +/- 0.69% and 33.6 +/- 0.13%) were lower (p < 0.001) than those obtained with control embryos (89.1 +/- 0.8% and 47.5 +/- 0.12%). Hatched VB embryos had a lower (p < 0.01) total cell number than hatched control embryos (70.3 +/- 4.5 versus 90.6 +/- 3.2, respectively). There was no difference between expanded VB and control blastocysts. In conclusion, blastocysts derived from in vitro culture of two-to-four cell pig embryos could be successfully vitrified using SOPS straws and Vit-Master (R). (c) 2007 Elsevier Inc. All rights reserved.