Minute changes to the culture environment of mouse pre-implantation embryos affect the health of the conceptus
Minute changes to the culture environment of mouse pre-implantation embryos affect the health of the conceptus
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DOI：
10.1016/j.apjr.2016.06.015
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发表时间：
2016-07
影响因子：
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通讯作者：
G. Koustas;C. Sjoblom
中科院分区：
文献类型：
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作者：
G. Koustas;C. Sjoblom
目的研究每天将胚胎从培养箱中取出并短时间暴露于 37.0 °C 的环境空气中对胚胎发育、基因表达和受孕健康的影响。方法使用 CBA F1 小鼠受精卵在其整个体外发育过程中每天暴露于 37.0 °C 的环境空气中 3、5 和 8 分钟进行体外横断面研究。对囊胚进行总细胞数、细胞凋亡和 H19 和 Igf2 表达分析，并将其转移至假孕受体女性体内，检查妊娠情况、胎儿和胎盘重量以及胎盘 H19 和 Igf2 表达。结果将植入前胚胎短暂暴露于 37.0 °C 的环境空气中，导致囊胚发育和总细胞数显着减少、凋亡细胞指数增加以及 H19 和 Igf2 受损。囊胚中 Igf2 的表达。怀孕第 18 天，着床率显着降低，胎儿和胎盘的重量显着减轻。胎盘中 H19 和 Igf2 表达水平显着降低。结论将小鼠植入前胚胎暴露在 37.0 °C 的环境空气中，即使是短暂的常规显微操作也不利于正常胚胎发育。我们的结果强调了培养环境的微小改变如何对胚胎的健康产生重大影响的重要性。
ObjectiveTo investigate the consequences of removing embryos from the incubator daily, and exposing them to ambient air at 37.0 °C for short periods of time, on embryo development, gene expression and the health of the conceptus.MethodsAnin vitrocross-sectional study was undertaken using CBA F1mouse zygotes exposed to ambient air at 37.0 °C for 3, 5, and 8 min daily during their entirein vitrodevelopment. Blastocysts were analyzed for total cell numbers, apoptosis, and H19and Igf2expression and transferred to pseudopregnant recipient females to examine pregnancy, fetal and placental weight, and placental H19and Igf2expression.ResultsShort exposure of pre-implantation embryos to ambient air at 37.0 °C resulted in a statistically significant reduction in blastocyst development and total cell numbers, increased apoptotic cell index, and impaired H19and Igf2expression in the blastocyst. Implantation rates were significantly reduced and fetuses and placentae displayed a significant reduced weight on day 18 of pregnancy. Placentae displayed significantly reduced levels of H19and Igf2expression.ConclusionsExposing mouse pre-implantation embryos to ambient air at 37.0 °C, even for brief periods for routine micromanipulations is detrimental to normal embryonic development. Our results highlight the importance of how small alterations in the culture environment can have major consequences for the health of the embryo.