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Merck
  • Generation and assessment of high-quality mouse oocytes and embryos following nicotinamide mononucleotide administration.

Generation and assessment of high-quality mouse oocytes and embryos following nicotinamide mononucleotide administration.

STAR protocols (2021-02-04)
Yilong Miao, Jingyue Chen, Qian Gao, Bo Xiong
要旨

The low quality of oocytes is one of the main causes of the suboptimal reproductive outcome of female mammals with advanced maternal age. Here, we present a detailed protocol to obtain high-quality oocytes and embryos from aged mice by nicotinamide mononucleotide (NMN) administration. We also describe fluorescence staining procedures to assess the organelle dynamics in oocytes, and in vitro fertilization and embryo culture systems to evaluate the influence of NMN on the fertilization ability and embryonic development potential. For complete information on the use and execution of this protocol, please refer to Miao et al. (2020).

材料
製品番号
ブランド
製品内容

Sigma-Aldrich
M2培地, With HEPES, without penicillin and streptomycin, liquid, sterile-filtered, suitable for mouse embryo cell culture
Sigma-Aldrich
β-ニコチンアミドモノヌクレオチド, ≥95% (HPLC)
Sigma-Aldrich
M16培地, With sodium bicarbonate and lactic acid, without penicillin and streptomycin, liquid, sterile-filtered
Sigma-Aldrich
タイロード溶液, 酸性, liquid, sterile-filtered, suitable for mouse embryo cell culture
Sigma-Aldrich
EmbryoMax® KSOM Mouse Embryo Media, (1X), Liquid, with 1/2 Amino Acids
Sigma-Aldrich
ウシ血清アルブミン ウシ血清由来, chromatographically purified, New Zealand origin, low endotoxin, suitable for cell culture, pH 7, ≥98%
Sigma-Aldrich
抗-α-チューブリン−FITC 抗体、マウスモノクローナル, clone DM1A, purified from hybridoma cell culture