Cumulus-oocyte complexes (COCs) were aspirated from antral follicles with a diameter of 3~8 mm using an 18 G needle attached to a 10-mL syringe. Only COCs with multiple layers of compacted cumulus cells were selected. These complexes were then washed three times and transferred to a 4-well dish (ThermoFisher Scientific, USA) containing 500 µL of an in vitro maturation (IVM) medium with gonadotropin including 10 IU/mL pregnant mare serum gonadotropin (PMSG; Intervet, The Netherlands) and 10 IU/mL human chorionic gonadotropin (hCG; Intervet). The cells were incubated at 39℃ with 5% CO2 in a humidified atmosphere [31 (link)]. After maturation for 22 h, the COCs were incubated in IVM medium without PMSG or hCG for an additional 22 h.
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In Vitro Maturation of Bovine Oocytes
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Antral follicles
Atmosphere
Cells
Cumulus cells
Dish
Gonadotropin
H hcg
Needle
Oocyte
Pregnant mare serum gonadotropin
Syringe
Corresponding Organization :
Other organizations : Animal and Plant Quarantine Agency, Kangwon National University, Seoul National University
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Variable analysis
independent variables
- Use of an 18 G needle attached to a 10-mL syringe to aspirate COCs from antral follicles with a diameter of 3-8 mm
- Selection of COCs with multiple layers of compacted cumulus cells
- Addition of PMSG (10 IU/mL) and hCG (10 IU/mL) to the IVM medium
- Maturation of COCs after incubation at 39°C with 5% CO2 in a humidified atmosphere
- Incubation temperature of 39°C
- Atmosphere with 5% CO2
- Humidified environment
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