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CD 4 cell isolation

Keith Bahjat Kbahjat at mindspring.com
Sat Sep 6 00:37:53 EST 1997

NO SPAM Mizuno wrote:

> I like the MACS system also.
> For our larger separations, we ended up buying the Vario
> MACS system,
> which costs quite a bit more than the starter MACS system.
> However, for separating large and for RARE cell
> populations, it works
> great.  I believe the Vario MACS is running about $5,000
> or so now.
> The conjugated "beads" (they really are not beads, and are
> smaller than your cells) cost about $400-500.
> The Vario MACS allows positive and negative selection
> capabilities,
> and with the use of cleaveable beads, there are many uses.
> We have isoalted human dendritic cells also with this
> system from
> peripheral blood.
> For CD4 cell isolation, I still recommend a FICOLL step
> prior to using
> the MACS system. I would also pass the cells through a
> nylon mesh to
> eliminate any cell clumps that may clog up the column.
> Hope this helps!
> -Mark Mizuno
> Scientist at Mizuno.com
> On Tue, 2 Sep 1997 23:15:00 -0400, "Scotty Wilcoxen"
> <wilcoxen at hsc.usc.edu> wrote:
> > In our lab we often need to make T cell rich
> populations, specifically CD4+
> >and CD8+, and we use the MACS system.  It gives great
> yields.  They use an
> >anti-CD4 or anti-CD8 Ab conjugated to a metallic bead.
> This solution is
> >then passed through a column with steel shavings in it.
> The column sits in
> >a huge magnet, and your desired fraction is either eluted
> out, or remains in
> >the column.   It is a bit slow, and VERY costly.  I hope
> this helps.
> >
> >Scott Wilcoxen
> >wilcoxen at hsc.usc.edu
> >
> >Robert Owe-Young wrote in article
> <33FC2D88.6B29 at cfi.unsw.edu.au>...
> >
> >>Does anyone have a favorite method for the isolation of
> large numbers of
> >>CD4 cells. We need to isolate CD4+ cells from apheresis
> specimens, and
> >>our current method has gone out of production.
> >>We routinely do a Ficoll step first
> >>I would like to hear from anyone who has experience with
> MACS, R&nce.D,
> >>or Baxter systems or any other similar commercially
> available system.
> >>
> >>thanks
> >>
> >>Dr Janet Mcpherson
> >>JJR Labs
> >>Sydney Australia
> >>
> >>Please reply to macpherson at angis.su.oz.au
> >
> >

 When you isolate CD4+ cells this way, why don't you get all
of the monocytes also, as they express CD4 (although
somewhat dimmer)?

Keith Bahjat
Graduate Assistant
University of Florida
College of Medicine
Gainesville, Florida
Kbahjat at mindspring.com

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