Mouse denuded oocyte in vitro maturation technology
An in vitro maturation, mouse technology, applied in the field of in vitro maturation of mouse nude eggs
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[0014] 1. Preparation of mature medium
[0015] TCM-199 culture medium (Gibco, Grand Island, NY, USA) add 1μg / ml 17β-E 2 , 24.2mg / l sodium pyruvate, 0.05IU / ml FSH, 0.05IU / ml LH and 10ng / ml EGF are made into mature culture medium. Prepare cysteamine and cystine into concentrated stock solutions of 20mmol / l and 100mmol / l respectively, store them at -20°C, dilute to the required concentration with mature culture broth before use (cysteamine: 100μmol / l; cysteamine) Acid: 200μmol / l). Since the commercial TCM-199 culture medium itself contains 83 μmol / l cystine, the final concentration of cystine used is 283 μmol / l.
[0016] 2. Preparation of mouse DO
[0017] Obtain ovaries from superovulated mice and puncture large follicles to obtain COC. Use a thin glass tube with a diameter slightly larger than that of the oocytes by pipetting repeatedly to remove the cumulus cells to obtain DO.
[0018] 3. Preparation of mouse or goat granule cell monolayer
[0019] Mouse or goat COC shed cumulus ...
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