I'm trying to reconstitute the immune system of irradiated Balb/c mice with LSK sorted cells, but they die every time. I tried to decrease the irradiation doses, they die later, but they still die. I suppose the cells I transfer are a bit damaged because of the sorting. Does anyone have an idea of what I should do?
No, but in the same time I reconstitute other mice with whole bone marrow, and it works, so I think it come from my sorted cells. Moreover, they are not all in the same cage, and they all die almost the same day.
In the first experiment they died at 7 days, I have been told it was because of the irradiation so I decreases doses and now they die at 10-15 days. First I tried 500 LSK cells, but then I also tried 10,000 cells (in papers they give 100 LSK, so it should work). I give C57B6 cells to Balb/c mice ( just while awaiting for CD45.1 Balb/c). Yes they are in clean conditions
Of course, I do not know the details of your experimental set-up, e.g. irradiation dose or number of effective stem cells for reconstitution. Perhaps the added paper using BM reconstitution after irradiation in mice will be helpful. Good luck with further studies !
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