Re: RE: Monocytes and Macs, AND MORE

From: Seth Ness (seth.ness@mssm.edu)
Date: Thu Dec 05 2002 - 20:12:09 EST


Hi,

>
>
> In our lab we routinely isolate monocytes from peripheral blood
(by
> Percoll). To put in my two cents, I will use the list of Mayumi to
> give my
> opinion and experience:
>
> - Percoll: http://memorias.ioc.fiocruz.br/952/3879.html
> In our hands the best solution. We perform this routinely and get
> puritiesaround 95% +/- 4%.
>

am i understanding correctly that this is actually a 3 step
procedure...

1. spin through lymphoprep
2. spin through percoll
3. one hour adherence

or is this wrong?

And here are some other possibilities, though i've never used
them...

1. centrifugal elutriation
2. a company names 'axis-shield' makes nycodenz and
omnipaque that can both be used in a single step isolation
procedure.
3. this reference that i'm not certain if its a one or two step
process...

J Immunol Methods 1991 Nov 22;144(2):247-51	Related Articles,
Links

A simple method for the purification of human peripheral blood
monocytes. A substitute for Sepracell-MN.
Denholm EM, Wolber FM.
Division of Hematology, Albany Medical College, NY 12208-3479.
Sepracell-MN has provided a simple, rapid means of isolating
peripheral blood monocytes. However this product is no longer
available. Consequentially we have developed a Percoll gradient
which matches Sepracell-MN in simplicity and yield of monocytes.
Using this Percoll gradient, an average of 7 x 10(6) monocytes with
a purity of 83% were obtained from 30-40 ml of blood. These
monocytes were at least 97% viable and responded to
chemotactic stimuli in comparable numbers to those prepared
using Sepracell-MN.
PMID: 1960422 [PubMed - indexed for MEDLINE]


---
Seth L. Ness M.D., Ph.D
Fellow in Human Genetics
Department of Human Genetics
Mount Sinai School of Medicine

Phone:212-241-6947
Fax:212-860-3316
sln8@columbia.edu



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