Hi everyone,
I have been trying to isolate CD3+ cells from mouse splenocytes using a nylon-wool column.
Although the viability of splenocytes isolated from balb/c mice is almost 98%, the cells rapidly die when they are passed through the nylon wool column.
I get almost 20-30% CD3+ cells out of the column with over 90% purity, but the viability evaluated just after elution is about 70% , and it drops to 50% after thawing the preserved cells.
Has anybody ever had this kind of problem with mouse T cells? Is the problem arising from nylon-wool itself, or is it normal for mouse T cells to die within such a short period of time?
(The whole protocol from spleen resection to T cell preservation takes about 6 hours for me using this protocol).
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