氧张力和湿度对体外受精小鼠胚胎植入前发育的影响:用延时摄影技术分析非加湿培养箱

Hiroyuki Watanabe, H. Ito, Ayumi Shintome, Hiroshi Suzuki
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摘要

为了研究氧张力和湿度对早期胚胎发育的影响,通过延时摄影评估体外受精产生的小鼠胚胎的植入前发育,以更高精度评估形态动力学发育。由ICR小鼠的精子和卵母细胞产生受精卵,并在KSOM中用延时摄影(CCM iBIS)在非加湿培养箱中在低氧或高压下培养。CCM iBIS在较低氧张力下胚胎发育到4细胞和胚泡阶段的比率显著高于CCM iBIS中在较高氧张力下的胚胎。受精96小时后,在低氧压力下培养的大量胚胎发育到孵化胚泡阶段。在较低的氧气压力下,胚胎发育更加同步。非湿润培养物不影响胚胎发育。平均而言,在低氧张力下培养的小鼠胚胎在受精后18小时达到2细胞,39小时达到3细胞,40小时达到4细胞,58小时开始压实,69小时达到桑椹胚,82小时达到胚泡。总之,较低的氧张力更好地支持体外受精小鼠胚胎的植入前发育,并且非湿润培养条件不影响体外胚胎发育。
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Effects of oxygen tension and humidity on the preimplantation development of mouse embryos produced by in vitro fertilization: analysis using a non-humidifying incubator with time-lapse cinematography
To examine the effects of oxygen tension and humidity on early embryonic development, the preimplantation development of mouse embryos produced by in vitro fertilization was assessed by time-lapse cinematography to evaluate morphokinetic development with higher precision. Zygotes were produced from spermatozoa and oocytes from ICR mice and cultured in KSOM under low or high oxygen tension in a non-humidified incubator with time-lapse cinematography (CCM-iBIS). The developmental rates of embryos to the 4-cell and blastocyst stages under lower oxygen tension in CCM-iBIS were significantly higher than those under higher oxygen tension in CCM-iBIS. Ninety-six hours after insemination, a large number of embryos cultured under low oxygen tension developed to the hatching blastocyst stage. Embryonic development was more synchronized under lower oxygen tension. Non-humidified cultures did not affect embryonic development. On average, mouse embryos cultured at lower oxygen tension reached 2-cell at 18 h, 3-cell at 39 h, 4-cell at 40 h, initiation of compaction at 58 h, morula at 69 h, and blastocyst at 82 h after insemination. In conclusion, lower oxygen tension better supports preimplantation development of mouse embryos fertilized in vitro, and non-humidified culture conditions do not influence the embryonic development in vitro.
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