This article describes a surgical technique for intra-uterine cell injection in immunocompromised nude mice. The method allows for the delivery of ovarian cancer cells into the uterine cavity, facilitating the study of tumor development and metastasis.
Intra-uterine cell injection allows the delivery of cells of interest into the uterine cavity of a recipient animal. To begin, prep an anesthetized immunocompromised nude mouse in the right lateral position. Make a surgical incision and dissect the abdominal muscle to expose its abdominal cavity.
Locate one of the uterine horns. These tubular structures are analogous to two distinct uteri, extending from the oviducts on both sides and opening into a single cervix.
Next, clamp the horn's anterior and posterior ends for segmentation. Now, slowly inject the ovarian cancer cell suspension into the segmented uterine lumen, ensuring efficient delivery of cells.
Subsequently, return the uterine horn into the abdominal cavity and suture the surgical incision. Allow the mouse to recover in a cage.
Within days of injection, tumors develop at the site of injection that metastasize to form secondary tumors in different tissues or organs in the peritoneal cavity.
To begin this procedure, anesthetize a 7 to 8 week old athymic nude mouse with 2% isoflurane. Then, verify that it is completely anesthetized by pinching its footpad. Next, place the animal on its right side on a sterile gauze pad with its head away from the experimenter. Apply ointment to both eyes to prevent dryness while under anesthesia.
Afterward, quickly insert the animal's head in a nose cone system connected to an isoflurane vaporizer. Disinfect the right side of the abdomen using alcohol pads followed by iodine. Then, place the sterile surgical drape over it. Using sterile surgical scissors and forceps, make a 1 to 2 centimeter skin incision on the lower left quadrant of the mouse.
After that, lift the muscle and make an incision in order to reach the peritoneum. Subsequently, dissect the oblique muscle to expose the abdominal cavity. Then, locate the left uterine horn. Using a sterile hemostat, clamp both the anterior and posterior sides of the horn.
Place the anterior clamp right below the fallopian tube and the posterior clamp right above the cervix. At this point, have the second experimenter inject the cell suspension by placing the cells containing needle at a 45-degree angle perpendicular to the horn. Then, slowly inject 50 microliters of cell suspension into the lumen of the uterine horn. Next, release the anterior clamp followed by the posterior clamp.
After that, place the uterine horn back in the abdominal cavity. Close the peritoneum using a synthetic absorbable suture. Then, close the skin using a tissue adhesive.
When finished, remove the mouse from the nose cone and place it back in its cage. Make sure the animal is awake and active before leaving it unattended. Provide ibuprofen in the drinking water for the first 48 hours after surgery.
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