I need to test the proliferation ability of anti-Her2 CAR T cells in response to Her2 expressing cell lines using 3H incorporation assay ,,, First I need to stop proliferation of my cancer cells using X ray ?
What is the irradiation dose will stop proliferation of cancer cell lines? doubling time 28hr
This is difficult to answer, since cell lines (also cancer cell lines) of different origins have different sensibilities to radiation. Easiest way would be to prepare a curve where you create a dose-response-curve and see at which dose your cells stop to proliferate (simple variant would be a growth curve).
Nevertheless, what is the question of your assay? With irradiation you may have unwanted side-effects, especially since you want to have a look on proliferation afterwards. When you use X-ray you induce a lot of damage (DNA-damage) resulting in cell cycle arrest (which you aim for) or exit of cell cycle (so-called proliferative cell death, which you certainly don't want). Could also go in the direction of apoptosis, depending on the sensitivity of your cells. The damage is not reversible in a classical sense.
If you just want to synchronize your cells, so that they start in G1 and go into S-Phase to analyze the effect on proliferation I would recommend a milder (reversible) treatment. There are a lots of proliferation inhibitors available that you could try. One example would be lovastatin (https://www.ncbi.nlm.nih.gov/pubmed/18677105). Some cells can be synchronized by just serum withdrawal and “started” again by changing into serum containing media. The damage/stress you induce with such methods is minimal compared to X-ray and would give you a better idea how the Her2-expression cells differs from your control.
In case you want to see the difference in proliferation dependent on irradiation I would recommend to perform the growth/clonogenic survival assay with your control cells first.
The purpose of the experiment and which cells are in which stage of the study will be a deciding factor. I support the answer by anja heselich.
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