side population wtih LSR/Novel Method

Lily Lai llai at ebioscience.com
Mon Mar 4 15:24:24 EST 2002


There is a simpler way to identify & enrich for the SP population.  A novel
mAb which binds to the SP stem cells  is now available in PE and biotin
conjugates.  Please see:
http://www.ebioscience.com/ebioscience/whatsnew/abcg2.html

You might also find the following references helpful:

Kim M, Tunquist H et al. 2002. The multidrug resistance transporter ABCG2
(breast cancer resistance protein 1) effluxes Hoechst 33342 and is
overexpressed in hematopoietic stem cells. Clin Cancer Res: 8(1):22-8.

Scharenber CW, Harkey MA. et al. 2002. The ABCG2 transporter is an efficient
Hoechst 33342 efflux pump and is preferentially expressed by immature human
hemtopoietic progenitors. Blood 15;99(2):507-12.

Zhou, S., J. D. Schuetz, et al. 2001. The ABC transporter Bcrp1/ABCG2 is
expressed in a wide variety of stem cells and is a molecular determinant of
the side-population phenotype. Nat Med 7(9): 1028-34.

Goodell MA, Rosenzweig M, et al. 1997. Dye efflux studies suggest that
hematopoietic stem cells expressing low or undetectable levels of CD34
antigen exist in multiple species. Nat Med 3(12):1337-45

Best regards,
Lily
www.ebioscience.com


----- Original Message -----
From: "Derek Davies" <daviesd2 at cancer.org.uk>
To: cyto-inbox
Sent: Thursday, February 28, 2002 2:08 AM
Subject: Re: side population wtih LSR


>
> Hi Kimmo,
>
> I think your problems are stemming (no pun intended, honest!) from the
> filter set-up of the LSR. If you look at the laser paths, the UV signals
> are directed to the FL4/5 PMTs via a 510LP filter so only fluorescence
> below 510nm will get to the detectors. As you say, this is a bit of a
> problem when looking for signals in the far red in the SP population.
> The problem will be worse if you alter the filter in front of FL4 to say
> a 650LP; keeping a 510/20 there will give you some signal but probably
> not far enough out in the red for the 'classical' SP population.
>
> There is no off the shelf solution as far as I can see other than
> modifying the LSR considerably - I believe BD may be working on this. It
> would be an adavnatge to be able to do SP analysis on a benchtop and I
> belive that the Cytomation CyAn may be better equipped to do this -
> currently we do all stem cell analysis on the MoFlo.
>
> Derek
>
> On Wed, 27 Feb 2002, Porkka Kimmo wrote:
> > Has anyone have experience on running a side population analysis of
human
> > bone marrow with the BD LSR? I'm using the Hoecst 33342 dye and a FITC
cell
> > surface marker and following the published protocol to the letter, but
still
> > cannot see any Hoechst fluorescence on the red channel. Filter setup?
> > Software compensation? Any help would be much appreciated.
>
> ************************************************************************
> Derek Davies                       Voice: (44) 020 7269 3394
> FACS Laboratory,                   FAX: (44) 020 7269 3100
> Cancer Research UK,                e_mail:derek.davies at cancer.org.uk
> London Research Institute,    mobile: 07790 604112
> 44 Lincolns Inn Fields,
> London, UK.
>
> Web Page: http://sci.cancerresearchuk.org/axp/facs/davies/index.html
>
> In tenebris lux
> *************************************************************************
>
>




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