To obtain fully-grown GV oocytes, 6–8 weeks of females were injected with 5 IU PMSG (Ningbo No.2 hormone factory, Zhejiang, China). After 44–48 h, GV oocytes were collected by puncturing the ovarian follicle and released with microcapillary pipettes in M2 media (M-7197, Sigma-Aldrich, Germany). For MII oocyte collection, mice were injected with PMSG as above and after 46–48 h with 5 IU of human chorionic gonadotropin (hCG) (Ningbo No.2 hormone factory, Zhejiang, China). MII oocytes were collected from the oviducts 14–16 h post hCG injection by digestion in M2 medium with hyaluronidase (MR-051-F, Millipore, USA). The detailed procedure to get meiotically incompetent PD5 and PD12 oocytes was described previously [25 (link), 62 (link)].
To obtain zygotes, female mice were mated with male mice with known fertility. Successful mating was confirmed by the presence of vaginal plugs. Zygotes were isolated from the oviduct of plugged females. The zygotes were cultured in KSOM medium (MR-107-D, Millipore, USA) and cultured in a 5% CO2 incubator.
To obtain zygotes, female mice were mated with male mice with known fertility. Successful mating was confirmed by the presence of vaginal plugs. Zygotes were isolated from the oviduct of plugged females. The zygotes were cultured in KSOM medium (MR-107-D, Millipore, USA) and cultured in a 5% CO2 incubator.
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