I want to keep WBCs (usually 2 to 3 days after collection).
After separation, cells were kept by mixing with RPMI, then the mixture was kept at 4°C.
After one day, all cells were dead. Normally, I use Histoprep (lymphocytes separation media) to separate WBCs from whole blood.
not more than 24 h in RPMI at 4ª, mainly for function tests. It is better to keep them frozen ( with a slow method!) at -120º if is going to be more than 24 h
Hi,
It dependent to the culture condition. Using recombinant cytokines you could able sustain them for more times. For example I kept purified lymphocyte of normal human in the presence of IL-2 for two weeks and differentiated dendritic cells with GM-CSF for more than one week.
I think it's better to keep them in serum in 4 degree. but 2-3 days is too long for RPMI. what do you want to do with WBCs? the answer will depend on it.
Thank you Abbas Ali Amini,
I need to separate WBC for Cross match (T-cell and B-cell test) to check matching in between patient and donor (In organ transplantation) actually,I need to store it for some days to check the viability and re-testing purpose.
If you store lymphocytes for 3 days, they will die. I think it's better to divide your cells into two parts: one for cross match (they must be fresh) and the other for viability. For viability, you must have a survival factor for cells; it could be a antigen or mitogen. What you are looking for from viability?
What is "re-testing'?
For the confirmation, some time we need to "re-testing",means to repeat the test. Where we want for re-sampling or wanting donor & patient blood's sample again.So,if WBC can be viable for 2 days then i don't need to call for re-sampling or again blood sample (taken) from donor and patient.
So ,to remove this i want to find a way to preserve the WBC alive for 1 more days to remove the re-sampling procedure.That's it.
if you want to repeat the test to confirm results, repeat it after 24 hours. by keeping cells in serum or RPMI-1640 containing 10% FBS in 4 degree, cells would be suitable enough to be used for other test.
since they will lose function, I do not recommend to keep cells for 2-3 days.
Thank you Abbas Ali Amini,hope it will work.I will try this method.Thank you again.