Neonatal Mouse Ovary Isolation: A Surgical Procedure to Harvest Pair of Ovaries from Neonatal Mouse Model

To isolate neonatal mouse ovaries, begin with a euthanized neonatal mouse pup in the supine position on a surgical platform. Make a long surgical incision on the skin and body wall to expose the abdominal cavity. Next, gently displace the intestines to reveal the underlying uterine horns.

The uterine horns are two long tubular structures that converge at the cervix and extend from the bladder towards the kidneys. Identify the ovaries - small, oval-shaped glands - located just below the kidneys at the tip of the uterine horns, connected via the oviducts.

Now, cut the attachment site of the ovary from each uterine horn to release the ovaries. Immediately transfer the excised ovaries into a sterile culture dish containing suitable pre-warmed media to maintain the viability of oocytes within the tissue. Place the culture dish onto the heated stage of a dissecting microscope.

Using a needle, carefully remove the ovarian bursa, a sac of connective tissue surrounding the ovary, and any remnants of the oviduct. Finally, transfer the ovaries into a multi-well plate containing media. The isolated ovaries can be used for downstream processing.