cryopreservation of lymphocytes - (Oct/21/2007 )
i have one Q
how much is the % of CD8 or CD4 from..1 ml of blood or PBMCs
the numbers of CD8 and CD4 will of course vary a lot between individuals, but in general:
CD8: 10-20% of PBMC
CD4 (witout CD14 which also express CD4): 20-40% of PBMC
check out separation of lymhocytes with Dynabeads from Dynal or Microbeads from Miltenyi..
As 10-30% of cells may die during freeze-thaw, you should count the live cells before stimulation with CD3/CD28 soyou make sure you add the correct amount of beads. Add IL-2 the second or third day.
We are really happy with Dynabeads for cell separations. They seem to be the most gentle method of cell isolation. Here is a useful page for choosing products for cell isolations: