Preparing cells for injection:
- Determine the total volume of injections (plan on 40 µl per gland plus enough for 1-2 extra glands per cell line/condition).
- Thaw out enough matrigel on ice for 50% of the total volume of injections.
- Trypsinize cells to a single cells suspension and count.
- Put 1 x 106 cells per gland plus enough for 1-2 extra glands per cell line/condition in a conical tube and pellet.
- Optional: wash 1X with PBS or media
- 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)
- 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.
- Keep cells in Matrigel:media mix on ice until injection.