Preparation of the Oocytes

1. Remove any residual cumulus cells from the oocytes by culturing in the hyaluronidase treatment medium (see Subheading 3.2., step 6) for up to 10 min in the hot box until the cumulus cells begin to loosen.

2. Pipet repeatedly with an automatic pipettor to remove the cumulus cells (see Note 11).

Fig. 2. Micromanipulation chamber with the holding pipet entering the chamber from the left side and the enucleation pipet entering from the right side.

3. Wash several times in wash medium to leave the cumulus cells behind.

4. Examine the oocytes under a dissecting microscope while turning the oocytes over to select mature oocytes with polar bodies.

5. Lift these oocytes out of the dish and store in the enucleation medium dish until required.

6. Take a manageable group of oocytes and place in the Hoechst/CB dish and culture for 15 min in the hot box.

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Diabetes is a disease that affects the way your body uses food. Normally, your body converts sugars, starches and other foods into a form of sugar called glucose. Your body uses glucose for fuel. The cells receive the glucose through the bloodstream. They then use insulin a hormone made by the pancreas to absorb the glucose, convert it into energy, and either use it or store it for later use. Learn more...

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