Vitrification, not cryoprotectant exposure, alters the expression of developmentally important genes in in vitro produced porcine blastocysts
Vitrification, not cryoprotectant exposure, alters the expression of developmentally important genes in in vitro produced porcine blastocysts
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DOI：
10.1016/j.cryobiol.2017.12.001
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发表时间：
2018-02-01
期刊：
影响因子：
2.7
通讯作者：
Sjoblom, Cecilia
中科院分区：
文献类型：
--
作者：
Bartolac, Louise K.;Lowe, Jenna L.;Sjoblom, Cecilia
胚胎的玻璃化冷冻是先进的牲畜育种计划和人类生育诊所的常见做法。最近的研究表明，玻璃化冷冻会导致一些胁迫相关基因的异常表达。然而，很少有研究检查玻璃化冷冻对发育重要基因的影响，并且没有在猪胚胎中进行过。本研究的目的是确定不同的玻璃化冷冻程序和冷冻保护剂组合对体外生产（IVP）猪囊胚中印迹基因表达的影响。玻璃化冷冻各组囊胚中胰岛素样生长因子2（IGF 2）的转录水平均低于非玻璃化冷冻对照组（P < 0.05）。暴露于冷冻保护剂的未玻璃化冷冻囊胚中IGF 2及其受体（IGF 2R）的表达水平与未玻璃化冷冻的对照囊胚相似（P > 0.05）。此外，使用乙二醇和丙二醇联合玻璃化冷冻的囊胚以及在封闭装置中玻璃化冷冻的囊胚的IGF2R转录水平与非玻璃化冷冻对照囊胚相似（P > 0.05）。总之，玻璃化冷冻，而不是暴露于冷冻保护剂，导致印迹基因IGF2和IGF2R的异常表达。研究发现，采用丙二醇或封闭装置的玻璃化冷冻方案对IVP猪囊胚中基因表达的破坏性最小。
The vitrification of embryos is common practice in advanced livestock breeding programs and in human fertility clinics. Recent studies have revealed that vitrification results in aberrant expression of a number of stress related genes. However, few studies have examined the effect that vitrification has on developmentally important genes, and none have been conducted in porcine embryos. The aim of this study was to determine the effects that different vitrification procedures and cryoprotectant combinations have on the expression of imprinted genes in in vitro produced (IVP) porcine blastocysts. The transcript levels of insulin-like growth factor 2 (IGF2) were lower in all groups of vitrified blastocysts compared to that in non-vitrified control blastocysts (P < 0.05). Expression levels of IGF2 and IGF2 receptor (IGF2R) in blastocysts that had been exposed to cryoprotectants without being vitrified were similar to that in non-vitrified control blastocysts (P > 0.05). Furthermore, blastocysts vitrified using ethylene glycol and propanediol combined, and those vitrified in a closed device, had IGF2R transcript levels similar to that in non-vitrified control blastocysts (P > 0.05). In conclusion, vitrification, but not exposure to cryoprotectants, caused aberrant expression of the imprinted genes IGF2 and IGF2R. Vitrification protocols that incorporated propanediol or a closed device were found to be least disruptive of gene expression in IVP porcine blastocysts.