Summary of neutrophil isolation methods

Alex Chang lpss at unixg.ubc.ca
Wed Mar 2 11:46:48 EST 1994


Hi, Folks:

Several days ago, I posted a message asking for efficient ways to
isolate neutrophils from human peripheral blood, I got lots of
helpful messages. I'd like to thank the people who responded to my
request.

Here is the summary of them.

A. Ficoll/Hypaque is the fastest way to isolate PMN.  There are
many formulations of ficoll/hypaque, some are much faster than
others. The formulation we use is homemade. It takes about one and
a half hours (one hour if you hurry).

Mike Herron<herro001 at maroon.tc.umn.edu>
Comment: not yet received the formulation yet.

B. We have used a one step method for neutrophil isolation that
works well for chemotaxis assays-Polymorphprep from Nycomed, used
to be distributed by Accurate, now by Gibco/BRL.

Cliff Beall <cbeall at magnus.acs.ohio-state.edu>
Research Scientist
Ohio State Biotech. Ctr.

C. Have you tried the Neutrophil Isolation medium. I have been
using it to separate human neutrophils for the past 4 years. It is
marketed by CARDINAL ASSOCIATES (Address: Cardinal Associates, P.O.
Box 5220 SANTA FE, NM 87501, Ross Robinson, 1-800-359-3249, fax
505-473-5637)
This one works perfectly well. You should keep it on the bench to
bring it to room temp before you start separating. It is a straight
forward procedure.

Padmakumar Narayanan <narayana at vet.vet.purdue.edu>
school of vet: medicine/purdue university

D.  I've been using magnetic beads (from Dynal) to separate rat B
spleenocytes from T spleenocytes. It is very efficient. I don't
known about neutrophils. It is an expensive method for start (the
magnetic separator is quit expensive comparing with the coated
magnetic beads), but at the end of the day, its worthwhile.
    I don't have the dynal catalogue here but if you need more
information, I can send to you latter.

Denise M. Moreira <D.Moreira at sheffield.ac.uk>

E. You should try a discontinous PERCOLL gradient of 55%/70%.
PERCOLL is available from Sigma and Pharmacia.
100% PERCOLL = stock solution of PERCOLL plus 0.9% w/v NaCl.

JOACHIM <nexejhey at rrzn-user.uni-hannover.de> 
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