Intra-uterine Cancer Cell Injection: A Technique to Deliver Cancer Cells for Establishing Intra-peritoneal Ovarian Cancer Mouse Model

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.

• Mouse Ovary – The egg-producing reproductive organ in female mice.
• Hemostat – A surgical tool used to control bleeding.
• Cell Injection – The process of inserting cells into a certain tissue or organ.
• Uterine Carcinoma – A type of cancer that affects the uterus.
• Oviducts - Tubes that link the ovaries to the uterus in female mammals.
• Uterine Horn - Part of the uterus in various animals, extending laterally to the Fallopian tubes.
• Forceps – A handheld hinged instrument used for grasping and holding objects.

• Introduce Mouse Ovary - Mouse ovaries produces eggs needed for reproduction (e.g., mouse ovary).
• Key Concepts - Summarizing principles like cell injection, and controlling bleeding (e.g., hemostat).
• Explain Underlying Mechanism - Brief overview of uterine carcinoma, a type of cancer affecting the uterus.
• Connect to Experiment - Link the background to the surgical study carried out on mice's ovary.

• What is cell injection and how is it done in mouse ovary?
• How are hemostats utilized in this surgical process?
• How does the uterine carcinoma affect mouse's reproductive system?

• Practical Applications – Mice surgeries provide vital information for human health (e.g., uterine carcinoma).
• Industry Impact - Could be useful to develop new treatments for humans (e.g., cell injection).
• Societal Importance - Aid understanding of human reproductive system (e.g., oviducts).
• Link to Scientific Advancements - Development in surgical methods and techniques for treatment (e.g., forceps).