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
> MUCH
> 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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