Xenograft Injection Prep ( Click to Download )

Preparing cells for injection:

1. Determine the total volume of injections (plan on 40 µl per gland plus enough for 1-2 extra glands per cell line/condition).
2. Thaw out enough matrigel on ice for 50% of the total volume of injections.
3. Trypsinize cells to a single cells suspension and count.
4. Put 1 x 10⁶ cells per gland plus enough for 1-2 extra glands per cell line/condition in a conical tube and pellet.
   1. Optional:  wash 1X with PBS or media
5. Make a solution of 50% Matrigel and 50% media, keep on ice (if cell pellet is going to take up significant volume, you could account for this by making 60% matrigel and 40% media)
6. Aspirate all media from pellets and resuspend in matrigel/media mix; use enough volume for 30-40 µl per injection.  Try to take into account the approximate volume of the pellet to the best of your ability to estimate when resuspending.
7. Keep cells in Matrigel:media mix on ice until injection.