Survival rate of frozen-thawed bovine IVF embryos in relation to exposure time using various cryoprotectants.
Survival rate of frozen-thawed bovine IVF embryos in relation to exposure time using various cryoprotectants.
复制标题
冻融牛 IVF 胚胎的存活率与使用各种冷冻保护剂的暴露时间的关系。
DOI：
10.1006/cryo.1993.1029
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发表时间：
1993
期刊：
影响因子：
2.7
通讯作者：
T. Suzuki
中科院分区：
文献类型：
--
作者：
M. Takagi;A. Boediono;S. Saha;T. Suzuki
研究了冷冻保护剂和暴露时间对体外受精胚胎冷冻解冻后体外受精胚胎发育的影响。用1.6M丙二醇(PG)、1.8M乙二醇(EG)、1.1M二甘醇和1.3M乙二醇单甲醚(EME)悬浮培养7~8天的囊胚和扩张囊胚。在实验一中，在暴露30分钟和120分钟后，将胚胎直接放入培养液中并清洗3次。胚胎与卵丘细胞在培养液中共培养。培养48h后，除PG的完全扩张率(55%比83%)外，所有4种冷冻保护剂在30min和120min的完全扩张率、孵化率和孵化率均无显著差异。在实验II中，为了比较四种冷冻保护剂的冷冻保护效果，将胚胎分别暴露于每种冷冻保护剂10~120min的不同时间，播种，以0.3℃/min的速度冷却到-30℃，然后投入液氮中储存，然后解冻。通过将胚胎与卵丘细胞在培养液中共培养来衡量存活率。PG(27-42%)和EG(37-56%)对胚胎孵化囊胚的发育无显著差异。EME暴露后胚胎发育率与暴露时间不超过40min的PG或EG相似(43-49%)。我们的发现表明，不同低温保护剂的毒性与暴露时间有关，PG和EG的毒性相对无毒，短期暴露于EME也是如此。
The relationship between cryoprotectant and exposure time on the development of IVF-derived embryos in culture after freezing and thawing was examined. Good to excellent quality Day 7 to 8 blastocysts and expanded blastocysts were suspended in 1.6 M propylene glycol (PG), 1.8 M ethylene glycol (EG), 1.1 M diethylene glycol, and 1.3 M ethylene glycol monomethyl ether (EME). In Experiment I, after 30 and 120 min of exposure, embryos were placed directly into culture medium and washed three times. Embryos were cocultured with cumulus cells in culture medium. After a 48-h culture, there were no significant differences in fully expanded, hatching, and hatched rates between the 30- and 120-min exposure to all four cryoprotectants except for the fully expanded rate with PG (55 vs 83%). In Experiment II, to compare the cryoprotective effects of the four cryoprotectants, embryos were exposed to each cryoprotectant for variable times from 10 to 120 min, seeded, and cooled to -30 degrees C at 0.3 degree C/min, plunged into liquid nitrogen for storage, and then thawed. Survival was measured by coculturing the embryos with cumulus cells in culture medium. There were no significant differences in development to hatched blastocysts of embryos exposed to either PG (27-42%) or EG (37-56%). The number of embryos developing after exposure to EME was similar to that in PG or EG when exposure time did not exceed 40 min (43-49%). Our findings suggest that the toxicity of different cryoprotectants is related to the exposure time and that PG and EG were relatively nontoxic as is exposure to EME for short periods.