Choi Eun Sol, Kawano Koga, Hiraya Misaki, Matsukawa Eibai, Yamada Masayasu
Laboratory of Reproductive Biology, Graduate School of Agriculture Kyoto University Kyoto Japan.
Nippon Medical & Chemical Instruments Co., LTD. Osaka Japan.
Reprod Med Biol. 2019 Jul 19;18(4):405-410. doi: 10.1002/rmb2.12288. eCollection 2019 Oct.
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.
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.
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%).
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.
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。
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。
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%)。
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。