Problem about T Cell Purification

NOSPAM Mizuno Mark at mizuno.com
Sun Jun 7 00:12:38 EST 1998


On 5 Jun 1998 06:19:29 GMT, boella at uxmail.ust.hk (CHAN Ella Wai Ching)
wrote:

> Dear all,
>
> I would like to know how to use percoll for T cell purification from 
> blood !!! 
>
> Regards,
>
> Ella

O.k... first of all, how restricted are you in your lab's budget?  I
would highly recommend using any of the cell separation bead systems
that are presently on the market right now.  They will save you time,
and if you wish to look at T Cell subsets, you will be able isolate
those even further.  The amount of time that you save is tremendous.
 I would run the blood through a ficoll (Lympholyte, or you can use
any other commercially available Ficoll variation) first.  This step
helps with your RBC and monocyte separation prior to using a column or
a bead system.  I haven't used percoll gradients in years due to the
time it takes to pour them and the fact that we needed to purify the T
Cells further.  Heck, I remember using Fetal Bovine Serum to separate
cells instead of Percoll or Ficoll, followed by Nylon Wool!

Bottom line, unless there is a real compelling reason why you need to
use Percoll, look for other methods.  Of course, keep in mind what
your final USE of your T Cells will be and choose a system
appropriately. Companies to consider will be Miltenyi Beads/columns (I
could NEVER get their name straight!), Cedar Lanes has a few Columns,
R & D Systems, etc... 

Have Fun...
-Mark Mizuno





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