This article describes a protocol for the transplantation of genetically modified human neural progenitor cells (NPCs) into an anesthetized mouse model. The procedure includes securing the mouse, preparing the surgical site, and ensuring proper injection techniques for optimal cell distribution.
Secure an anesthetized mouse in a stereotaxic frame.
Apply eye ointment to prevent corneal drying.
Shave the scalp and disinfect the skin.
Insert the ear bars to stabilize the skull.
Then, make a midline incision to expose the skull.
Drill a hole through the skull at the target coordinate.
Load a syringe with genetically modified human neural progenitor cells or NPCs expressing luciferase and a fluorescent protein for visualization.
Align the syringe with the target site and lower the needle to the desired depth.
Apply tissue adhesive around the needle to prevent cell leakage.
Inject the cells slowly to ensure uniform distribution.
Hold the needle in place briefly to prevent backflow of the cell suspension.
Finally, withdraw the needle slowly and close the incision.
The mouse model with transplanted NPCs is now ready for visualization.
Transport the animal from the induction chamber to the stereotaxic frame, maintaining anesthesia using a face mask.
Apply ophthalmic lubricant to prevent the eyes from drying out. Shave the mouse scalp with an electric razor and disinfect the skin with 5 percent betadine solution using cotton swabs. Secure the mouse head and insert the ear bars into the external meatus.
Drill a hole with a diameter of two to three millimeters through the skull with a surgical dental drill. Resuspend the prepared cells in the tube and draw two microliters of cell suspension into a syringe. Place the syringe above the target site and slowly move the needle to the surface of the dura and calculate the depth coordinates.
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