Both maturation and early embryonic development rates of GV-stage oocytes were significantly higher (P < 0.01) in the IVM-medium (maturation: 75.7%; blastocyst: 12.9%) compared with control (maturation: 55.7%; blastocyst: 0.0%).
The Es3L (lower sub-member of the third member of the Eocene Shahejie Formation) shale of Jiyang depression was a set of relatively thick and widely deposited lacustrine sediments with high organic richness and medium thermal maturation (0.7%≤Ro≤1.1%), which rapidly emerges an important and target shale-oil play in Eastern China.
The transfected DCs were then cultured 2 days in medium containing maturation stimulants, MCM and poly [I] : poly [C].
Pooled oocytes were matured in maturation medium for 24 h in 200 µl within a Nunc 4-well dish (Nunclon, Roskilde, Denmark), then inseminated in BIVF medium (200 µl, 2 × 10 / ml final sperm concentration).
The present experiments compared the maturational and developmental competence of immature oocytes derived from stimulated cycles, following culture in a newly designed in-vitro maturation medium (IVM-medium) or in standard tissue culture medium (TCM-199; control).
Globular/heart shaped embryos developed and produced torpedo and cotyledon stage embryos upon subculturing on embryo maturation medium EM8 (medium containing MS salts, B5 vitamins, 0.5 μM BA, and 2% sucrose).
As a control, we used COCs matured only in maturation medium and cocultured with granulosa cells [ 18].
Briefly, the maturation medium consisted of TCM 199 medium with Earle's salt and 25 mM sodium bicarbonate, supplemented with 10% oestrous cow serum, 1 mM L-glutamine, 0.2 mM sodium pyruvate, 1.4 IU/ml pregnant mare serum gonadotropin and 0.7 IU/ml human chorionic gonadotropin (Suigonan Vet, Intervet Scandinavia, Skovlunde, Denmark), 50 ng/ml epidermal growth factor (EGF) and 50 μg gentamycin.
To develop and test a simple, feasible approach to improve spindle assembly and developmental competence of human in vitro matured oocytes by parthenogenetic activation in maturation medium.
Concisely, prior to the SCNT procedure, cumulus-denuded in vitro matured gilt/sow oocytes were incubated in the maturation medium supplemented with 0.4 μg/mL demecolcine (DMCC) for 50 to 60 min.
Groups of ten COCs with homogenous cytoplasm and more than three layers of compact cumulus cells were placed in 50-μl drops of maturation medium (MM) under mineral oil and matured for 24 h at 39 °C under an atmosphere of 6%% CO2 in air with maximum humidity.
