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All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. MN-anti-miR10b administration
NOTE: In this study, the injections were done once a week for 6 weeks beginning 7 days after implantation, but the same protocol can be used for various injection frequencies.
- Weigh the mouse and calculate the volume of MN (magnetic nanoparticle)-anti-miR10b to administer to the mouse. Doses of MN-anti-miR10b are given at 20 mg Fe (iron)/kg in murine GBM (glioblastoma multiforme) models. For example, a 20 g mouse receives an 80 µL dosage of 5 mg Fe/mL MN-anti-miR10b. Control mice receive an equal volume of PBS (phosphate buffered saline) consistent with this dosage scheme.
- Prepare a 28G syringe with the dose of MN-anti-miR10b and ensure there are no bubbles in the injection solution.
- Anesthetize the mouse under 2%-4% isoflurane for induction and 1.5%-2% for maintenance with a 1.5 L/min flow rate in an induction box. Move the mouse to a nose cone and continue to administer anesthesia. Check the depth of anesthesia and apply vet ointment.
- Sterilize the tail with a 70% isopropanol wipe and allow excess alcohol to dry.
- Rotate the tail to position the lateral caudal veins at the top of the tail. Then, aiming for the vein, insert the needle into the tail and establish a vacuum by pulling back the plunger.
- Insert the needle until blood enters the syringe, indicating successful entry into the vein.
- Slowly inject MN-anti-miR10b and retract the needle after complete delivery. Apply pressure to the injection site with gauze until the bleeding stops.
- Remove the mouse from the nose cone. Monitor the mouse until fully recovered and ambulatory.
NOTE: Bolus administration of nanoparticles may cause respiratory distress. Carefully monitor the mouse and immediately apply light chest compressions if there are signs of respiratory distress.