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How to implant tumors?

How to implant tumors? Orthotopic tumor implantation into 4th mammary gland in mice Materials: • Cultured tumor cells • 50 mL Conical Tubes • Sterile 27-30 G needles • 1 mL syringes • Isoflurane • Wet ice • 70% EtOH In this video syngeneic orthotopic 2e5 4T1 breast tumor model in BalB/c mice was used as an example. 1. Draw cells without needle attached. 2. Anesthetize mice with isoflurane. 3. Mix tumor cells suspension well but gentle. Draw cells without needle attached. 4. Identify abdominal mammary gland #4. Spray injection site with 70% EtOH. 5. Lift up the skin close to the 4th mammary gland nipple area with the forceps prior to injecting. Inject 100 uL of the tumor cell suspension. Needle goes under the nipple area. If properly injected into mammary fat pad, you will see a visible bump and no leakage of the cells in the injection site. 6. Place the animal back in its cage. Observe the mice two times a week for tumor development and growth. Tumors will become palpable in about 5 days following injection, and you can start measuring them by calipers. For the tumor calipering procedure check the link --    • How to measure tumors?  .