Universitary Institute IVI Valencia, Valencia, Spain.
J Assist Reprod Genet. 2013 Aug;30(8):1043-7. doi: 10.1007/s10815-013-0036-8. Epub 2013 Jul 3.
To determine the survival and subsequent in vitro development of human cleavage stage embryos and hatched blastocysts following varying periods of short-term storage at 4 °C, using tripronucleated human embryos (TPN) as a model.
TPN cleavage embryos and hatched blastocysts short-term stored at 4 °C for 0 h (control), 24 h and 48 h. The main outcome measures were: survival rates (SR) and in vitro developmental ability (blastocyst rate and blastocyst-re-expansion rate) in each of the groups after storage.
Cleavage-stage TPN survived at comparable rates to controls, regardless of storage time (average: 97.3 %). The in vitro development of cleavage-stage TPN stored for 24 h was comparable to that of controls (average 64.7 %), but was significantly impaired when storage lasted 48-h (20.8 %). After artificial shrinkage, SR was comparable in 24-h-stored and non-stored hatched blastocysts (85.7 %; p > 0.05), but was significantly impaired in the 48-h-stored group (20.0 %). Following 24-h storage, the re-expansion rate of hatched blastocysts was similar to that of controls (average: 57.1 %; p > 0.05), but was higher than that of the 48-h-stored group (15.0 %; p < 0.05).
TPN human cleavage embryos and blastocysts can be successfully stored short-term for up to 24 h at 4 °C without using cryoprotectants without any significant negative impact on survival or subsequent in vitro development.
以三原核(TPN)人类胚胎为模型,确定人类卵裂期胚胎和孵育囊胚在 4°C 短期储存不同时间后的存活率和随后的体外发育情况。
将 TPN 卵裂胚胎和孵育囊胚在 4°C 下分别短期储存 0 h(对照组)、24 h 和 48 h。主要观察指标为:各组储存后胚胎的存活率(SR)和体外发育能力(囊胚率和囊胚再扩张率)。
无论储存时间如何(平均:97.3%),卵裂期 TPN 的存活率与对照组相当。24 h 储存的卵裂期 TPN 的体外发育与对照组相当(平均 64.7%),但当储存时间延长至 48 h 时,其发育明显受损(20.8%)。经人工收缩后,24 h 储存和未储存的孵育囊胚的 SR 相当(85.7%;p>0.05),但 48 h 储存组的 SR 明显受损(20.0%)。24 h 储存后,孵育囊胚的再扩张率与对照组相似(平均:57.1%;p>0.05),但高于 48 h 储存组(15.0%;p<0.05)。
无需使用冷冻保护剂,TPN 人类卵裂期胚胎和囊胚可在 4°C 下成功短期储存长达 24 h,对存活率和随后的体外发育无明显负面影响。