Monocyte Isolation ?

u70z1da at sun1.lrz-muenchen.de u70z1da at sun1.lrz-muenchen.de
Mon Oct 9 13:42:37 EST 1995


I am currently trying to separate monocytes from peripheral human venous
blood. For this purpose I am using Optiprep (Iodixanol) (available through
Nycomed). 
In doing this I was succsessful only once, though the monocytes were 
heavily contaminated with thrombocytes. I was however not able to reproduce
this result.
The Nycomed hotline has no idea what went wrong. I am strictly following the
manual.

The problems I encountered so far were mainly:

1. After the first centrifugation I get two bands of cells instead of one as
   it should be and after the second centrifugation I get no monocyte band.

2. Alternatively I get white clumps of thrombocytes/monocytes.

What I`ve tried so far to solve the problem is to add more EDTA than usual to
avoid 2. This seemed to have a succsess.
I took the blood from different donors, so I don`t think that this is a reason
for the failure.

If anybody has any ideas I would be very thankful for a reply.

Bjoern Lorenzen
(u70z1da at sunserver.lrz-muenchen.de)
Munich, Germany




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