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单独或联合使用丙酮酸和二甲基-α-酮戊二酸对体外培养的小鼠受精卵植入前和植入后发育的影响。

Effects of pyruvate and dimethyl-α-ketoglutarate, either alone or in combination, on pre- and post-implantation development of mouse zygotes cultured in vitro.

作者信息

Choi Eun Sol, Kawano Koga, Hiraya Misaki, Matsukawa Eibai, Yamada Masayasu

机构信息

Laboratory of Reproductive Biology, Graduate School of Agriculture Kyoto University Kyoto Japan.

Nippon Medical & Chemical Instruments Co., LTD. Osaka Japan.

出版信息

Reprod Med Biol. 2019 Jul 19;18(4):405-410. doi: 10.1002/rmb2.12288. eCollection 2019 Oct.

Abstract

PURPOSE

Dimethyl α-ketoglutarate (dm-α-KG) promotes in vitro development to blastocysts of C57BL/6J X C3He F1 mouse zygotes cultured in medium lacking pyruvate. Here, we examined the effects of pyruvate and dm-α-KG on in vitro development to blastocysts of ICR mouse zygotes and their post-implantation developmental ability.

METHODS

Zygotes were cultured in medium with pyruvate at 0-0.2 mmol/L in the presence or absence of 1 mmol/L dm-α-KG for 96 hours and evaluated for blastocyst formation rates. The resultant blastocysts were non-surgically transferred to surrogates and evaluated for birth rates.

RESULTS

In medium lacking pyruvate, zygotes could not develop beyond the two-cell stage, in the presence or absence of dm-α-KG. However, the blastocyst formation rate in medium with 0.01 mmol/L pyruvate (12%) was markedly increased with addition of dm-α-KG (49%). Around 80% of embryos developed to blastocysts in medium with 0.2 mmol/L pyruvate, in the presence or absence of dm-α-KG. Importantly, birth rate was markedly improved by treatment with 0.2 mmol/L pyruvate and dm-αKG (31.0%), compared with those with pyruvate treatment alone (16.3%).

CONCLUSIONS

Pyruvate and dm-α-KG synergistically work during in vitro culture to markedly improve the blastocyst formation rate and post-implantation developmental ability of the resultant blastocysts in ICR mice.

摘要

目的

α-酮戊二酸二甲酯(dm-α-KG)可促进在缺乏丙酮酸的培养基中培养的C57BL/6J×C3He F1小鼠受精卵体外发育至囊胚。在此,我们研究了丙酮酸和dm-α-KG对ICR小鼠受精卵体外发育至囊胚及其着床后发育能力的影响。

方法

将受精卵在含有0-0.2 mmol/L丙酮酸的培养基中,在有或无1 mmol/L dm-α-KG的情况下培养96小时,并评估囊胚形成率。将所得囊胚通过非手术方式移植到代孕母鼠体内,并评估出生率。

结果

在缺乏丙酮酸的培养基中,无论有无dm-α-KG,受精卵都无法发育到二细胞期以后。然而,在含有0.01 mmol/L丙酮酸的培养基中,添加dm-α-KG(49%)后,囊胚形成率(12%)显著提高。在含有0.2 mmol/L丙酮酸的培养基中,无论有无dm-α-KG,约80%的胚胎发育到囊胚。重要的是,与单独使用丙酮酸处理(16.3%)相比,用0.2 mmol/L丙酮酸和dm-α-KG处理后的出生率显著提高(31.0%)。

结论

丙酮酸和dm-α-KG在体外培养过程中协同作用,显著提高ICR小鼠所得囊胚的囊胚形成率和着床后发育能力。

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