For as long as we can remember we have requested lithium heparin be used as the anti-coagulant from our buffy coat suppliers. This was largely due to a former lab head who insisted this was the case. Does anyone have an opinion on whether EDTA would be more appropriate as our supplier is suggesting we try this as we don't receive the buffy coats until up to 24 hours after collection? Would EDTA help us increase our yield of PBMC?
Sometimes it also depends on the temperature and the spin condition. We centrifuged at 4 degree celsius, Acc -4, Deacc - 1, 1800 rpm, it worked well.
EDTA or heparin also depends on your population of interest. EDTA is good for plasma analysis.
We are generally looking at CD8 depleted PBMC, or isolated CD4 cells or monocytes.
We have been using EDTA-Vacutainers with great success for monocytes from PBMCs. However, 24 hours is a long time to wait once blood has been taken from a patient?
They are healthy donors and we can only stipulate that the blood has been drawn in the previous 24 hours, but it is often quite clear we are at the limits of this when we process the buffy coats based on the recovery of PBMC. Thanks for your help.
- Badges
- Science topic
- Similar topics
- Biological Science
- Biochemistry
- Enzymology
- Enzymes