Porcine Cumulus Oocyte Complex Isolation: A Technique to Isolate Cumulus Oocyte Complex from Porcine Ovarian Follicle for In Vitro Fertilization

To isolate the COCs from medium-sized follicles, transfer two to three ovaries to a sterile 10-cm Petri dish filled with HM. Gently cut the surface of the protruding ovarian follicles with a sterile surgical # 15 blade, which will cause the follicular fluid, and the COCs to flow out into the Petri dish. Aspirate the follicular contents with a 28-gauge needle attached to a disposable syringe, and transfer it into another Petri dish.

To prepare the IVF Petri dishes, add 1 milliliter of HM to the central wells, and place three to four drops of HM in the outer rings. Then, use a polycarbonate micropipette to move the undamaged COCs to drops of HM in the outer rings, and briefly rinse them three to four times. When finished, individually transfer them into the central well. Store the IVF plates in the incubator.