Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Peking Union Medical College, Beijing, People's Republic of China.
Fertil Steril. 2010 May 15;93(8):2729-33. doi: 10.1016/j.fertnstert.2010.03.025. Epub 2010 Apr 18.
To investigate the effects of embryo vitrification on the methylation status of the H19/Igf2 differentially methylated domain (DMD) and the expression of those genes.
Comparative and controlled study.
A university-affiliated hospital.
ICR female and male mice.
INTERVENTION(S): Day 14 fetuses that arose from copulation of male and female mice were taken as the control group. Embryos resulting from superovulation, IVF, in vitro culture, and embryo transfer were taken as the IVF group. Embryos that were vitrified were taken as the vitrified group.
MAIN OUTCOME MEASURE(S): The methylation status of H19/Igf2 DMD was analyzed by cloning and sequencing following DNA bisulfite treatment. The expression of H19 and Igf2 was quantified by real-time reverse-transcription polymerase chain reaction.
RESULT(S): Loss of methylation was found in the H19/Igf2 DMD of the fetuses from the IVF group, but it was more severe in the vitrified group. H19 expression was significantly increased in the IVF and vitrified groups, as compared with the control group; however, the quantity in the vitrified group was less than that detected in the IVF group. Igf2 expression in the IVF and vitrified groups was found to have decreased significantly, whereas the Igf2 expression in the vitrified group was greater than in the IVF group.
Embryo vitrification aggravated the loss of methylation in the H19/Igf2 DMD, and compensated for the perturbed expression of H19 by IVF procedures.
研究胚胎玻璃化冷冻对 H19/Igf2 差异甲基化区（DMD）甲基化状态及基因表达的影响。
对比对照研究。
一所大学附属医院。
ICR 雌性和雄性小鼠。
将交配后第 14 天的胎鼠作为对照组。将超排卵、体外受精（IVF）、体外培养和胚胎移植获得的胚胎作为 IVF 组。玻璃化冷冻的胚胎作为玻璃化组。
DNA 亚硫酸氢盐处理后克隆测序分析 H19/Igf2 DMD 的甲基化状态，实时逆转录聚合酶链反应定量分析 H19 和 Igf2 的表达。
IVF 组胎鼠 H19/Igf2 DMD 出现去甲基化，但玻璃化组更为严重。IVF 和玻璃化组 H19 表达均明显高于对照组，但玻璃化组的量少于 IVF 组。IVF 和玻璃化组 Igf2 表达明显降低，而玻璃化组 Igf2 表达高于 IVF 组。
胚胎玻璃化冷冻加重了 H19/Igf2 DMD 的去甲基化，通过 IVF 程序补偿了 H19 表达的紊乱。
