Hi,
We have been struggling to establish the A20 (ATCC TIB-208) mouse B-cell lymphoma subcutaneously injected into the right flank. However, in our case, this model varies a lot between separate experiments in terms of engraftment.
We have tried different doses of A20 cells injected in PBS or Matrigel (1:1), with or without preconditioning (3 Gy/5 Gy TBI/100 mg/kg cyclophosphamide). It turned out we achieved successful engraftment with cells injected in Matrigel, with and without preconditioning; hence, we decided to proceed without lymphodepletion. We use 2 million cells per flank injected in 100 μl Matrigel (CAT 354234).
Unfortunately, we encountered another issue: in some experiments, we achieved a 100% engraftment rate, while in the rest of the cases, we observed the tumors growing until they reach ~50-80 mm3 (around day 10 post-inoculation), then they start to shrink rapidly, and in a few days, the mice are completely tumor-free. We use cells from the same batch (purchased from ATCC) and thaw fresh cells before injections. We do not use Fluc+ cells.
The last time we harvested the tumor after it started to decrease in size and processed it to make a single-cell suspension, we stained it for flow, and we saw it was heavily infiltrated with T cells suggesting that the host immune system rejected the tumor, even though this model is syngeneic and was injected to the same strain of mice.
Does anyone have advice on how to set up this model, please?
Best, Patrycja
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