Recently, I am doing the xenograft tumor growth assay. I am supposed to inject same cell numbers in different groups. However, I found cells in some groups aggregate quickly, which may make them less viable. Are there some solutions to compensate the effects caused by cell number difference?
1. Count and prepare the cell suspension in incomplete media.
2. Keep the cells at 4 degree Celsius all the time and do the process faster.
3. Mix your cell suspension gently before every injection.
1. Tapping the syringe before injections can help to reduce aggregates clogging the needle (point 3 of Swarnendra). Clogging the needles might be the reason to get less or no tumors (as you had noted as less viable; in such clogged occasions only the liquid pass through the needle not the cells).
2. Keeping cells in 4 degree/ice might reduce tumorigenecity in a cell type dependent manner.
If you are doing subcutaneous or mammary fat pad injection it might be worth mixing them 1:1 in Matrigel (keep at 4 degrees). This is quite viscous and prevents cells from clumping and also from leaking out of any injection site.
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