- 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
PMID33532742
RESUMEN
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).
MATERIALES
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Sigma-Aldrich
Medio M2, With HEPES, without penicillin and streptomycin, liquid, sterile-filtered, suitable for mouse embryo cell culture
Sigma-Aldrich
Tyrode′s Solution, Acidic, liquid, sterile-filtered, suitable for mouse embryo cell culture
Sigma-Aldrich
M16 Medium, With sodium bicarbonate and lactic acid, without penicillin and streptomycin, liquid, sterile-filtered
Sigma-Aldrich
Seroalbúmina bovina, chromatographically purified, New Zealand origin, low endotoxin, suitable for cell culture, pH 7, ≥98%