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首页> 外文期刊>Cryobiology: International Journal of Low Temperature Biology and Medicine >Cryopreservation of bovine in Vitro produced embryos using ethylene glycol in controlled freezing or vitrification
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Cryopreservation of bovine in Vitro produced embryos using ethylene glycol in controlled freezing or vitrification

机译：在受控冷冻或玻璃化中使用乙二醇对牛体外产生的胚胎进行超低温保存

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In this study, the cryoprotectant ethylene glycol (EG) was tested for its ability to improve and facilitate the cryopreservation of in vitro produced (IVP) bovine embryos. Embryos were cryopreserved in EG solutions supplemented with either newborn calf serum (NBCS) or polyvinyl alcohol (PVA). To assess EG toxicity, the embryos were equilibrated in EG concentrations from 1.8 to 8.9 M at room temperature for 10 min and then cultured for 72 h on a cumulus cell monolayer. The hatching rate was highest for day 7 blastocysts frozen in 3.6 MEG (98%) and was not different from the control group (85%). The controlled freezing (0.3 deg C/min to -35 deg C) of expanded day 7 blastocysts resulted in a hatching rate of 81%. which was similar to that of the honfrozen controls (76%). Differential staining revealed only very few degenerate blastomeres attributed to freezing and thawing. Upon direct nonsurgical transfer of day 7 expanded blastocysts frozen in 3.6 MEG. a pregnancy rate of 43% was achieved, while the pregnancy rate after transfer of other developmental stages was significantly lower (22% with expanded day 8 blastocysts). When bovine IVP embryos were incubated at room temperature in 7.2 MEG preceded by preequilibration in 3.6 MEG. the hatching rate of day 7 expanded blastocysts reached 93%. Upon vitrification of IVP day 7 and day 8 blastocysts and expanded blastocysts in 7.2 MEG. the latter showed a higher hatching rate (42%) than blastocysts (12%) Overall. PVA as supplement to the basic freezing solution instead of NBCS had deleterious effects on survival after controlled freezing or vitrification. The simple cryopreservation protocol employed in this study and the low toxicity of ethylene glycol highlight the usefulness of this approach for controlled freezing of IVP embryos. However. further experiments are needed to improve the pregnancy rate following embryo transfer and to enhance survival after vitrification.
机译：在这项研究中，测试了冷冻保护剂乙二醇（EG）改善和促进体外生产（IVP）牛胚胎冷冻保存的能力。将胚胎冷冻在补充有新生小牛血清（NBCS）或聚乙烯醇（PVA）的EG溶液中。为了评估EG毒性，在室温下将胚胎在EG浓度从1.8到8.9 M平衡10分钟，然后在单层卵丘细胞上培养72 h。冷冻在3.6 MEG中的第7天囊胚的孵化率最高（98％），与对照组无差异（85％）。扩张的第7天胚泡的受控冻结（0.3℃/ min至-35℃）导致孵化率为81％。与仿制控件（76％）相似。差异染色显示仅极少数归因于冷冻和解冻的简并卵裂球。在第7天直接非手术转移后，将膨胀的胚泡冷冻在3.6 MEG中。妊娠率达到43％，而其他发育阶段转移后的妊娠率则明显降低（第8天囊胚扩大时为22％）。当牛IVP胚胎在室温下于7.2 MEG中孵育，然后在3.6 MEG中进行预平衡。第7天膨胀胚泡的孵化率达到93％。在IVP第7天和第8天囊胚和7.2 MEG中扩展的囊胚玻璃化后。后者的孵化率（42％）高于胚泡​​（12％）。 PVA代替NBCS作为基本冷冻溶液的补充，对受控冷冻或玻璃化后的存活具有有害影响。在这项研究中使用的简单的冷冻保存协议和乙二醇的低毒性突出了这种方法对IVP胚胎控制冷冻的有用性。然而。需要进一步的实验来提高胚胎移植后的怀孕率并提高玻璃化后的存活率。

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