This article outlines the procedure for isolating neonatal mouse ovaries for further research. It details the surgical techniques required to extract the ovaries while maintaining their viability for downstream applications.
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.
Begin by transferring 1 milliliter of dissection medium into each sterile glass embryo dish. After culling the 0 to 5 postnatal day old pups, grasp the skin covering the abdominal wall with fine forceps and incise the skin and body wall. Pull open this large incision, exposing the abdomen. The bladder is usually engorged at this stage and can be punctured to make dissection easier.
Using watchmaker forceps, move aside the guts. Then, on either side, follow the uterine horns from the bladder up to the kidney. The ovary is a cloud-like structure located just below the kidney at the top of the uterus. Grasp the ovary gently and, using scissors, cut it free from the uterus. After dissecting out both ovaries, transfer them to dishes filled with warm dissection medium.
Use a dissection microscope with a stage heated at 37 degrees Celsius to dissect the ovaries. Using insulin needles, trim away the bursa sac and any excess material including the fallopian tube until only the ovary remains. Then, transfer each ovary into the well of a prepared plate using a curved pipette.
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