[Cytometry] Help with macrophage detachment

Nebe-Von-Caron, G g.nebe-von-caron at spdspark.com
Fri Feb 6 11:27:27 EST 2009


Has anyone tried to grow the macrophages on cygel? If I understand it
right it is normally used to trap nonadherent cells but as it dissolves
when cooled it might release cells that grow on top of it as well.


Gerhard Nebe-von-Caron 
Research Scientist and Biomedical Engineer 
SPDspark, www.swissprecisiondiagnostics.com
SPD Development Company Limited
Priory Business Park
Stannard Way
Bedford    MK44 3UP
 
Company Registration No.  6032177
Mob+44(0)7792-116609
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-----Original Message-----
From: cytometry-bounces at lists.purdue.edu
[mailto:cytometry-bounces at lists.purdue.edu] On Behalf Of Paul W. Price
Sent: 05 February 2009 18:45
To: Parlane,Natalie; 'Wiener,Doris'; cytometry at lists.purdue.edu
Subject: Re: [Cytometry] Help with macrophage detachment

Microbiology Petri Dishes are very good, and so are the T flasks
designed for suspension cultures (Sarstedt among others).  Having still
observed significant numbers of adherent cells using both, I found that
one or two rounds of ice-cold PBS (Ca++ and Mg++ free) containing 0.2mM
EDTA for 5 minutes improves recovery.

Paul 

Paul W. Price, Ph.D.
Director of R&D
Abeome Corporation
www.abeomecorp.com
Tel. 706-542-7889

-----Original Message-----
>From: "Parlane, Natalie" <natalie.parlane at agresearch.co.nz>
>Sent: Feb 3, 2009 7:30 PM
>To: "'Wiener, Doris'" <dwiener at health.usf.edu>, 
>"cytometry at lists.purdue.edu" <cytometry at lists.purdue.edu>
>Subject: Re: [Cytometry] Help with macrophage detachment
>
>Have you tried culturing in petri dishes (you know the common round (or
square) ones used for microbiology culture)? They are sterile, the MACs
attach but then easily come off easily with a gentle tap. It was a long
time ago but you may need bring to room temp prior, too.  
>I believe the other tissue culture plates are coated to aid attachment.
>Good luck
>Natalie
>
>
>Natalie Parlane
>Research associate
>Infectious diseases
>Hopkirk Research Institute
>AgResearch
>New Zealand
>T +64 6 351 8692
>F +64 6 353 7853
>E natalie.parlane at agresearch.co.nz
>
>
>-----Original Message-----
>From: cytometry-bounces at lists.purdue.edu 
>[mailto:cytometry-bounces at lists.purdue.edu] On Behalf Of Wiener, Doris
>Sent: Saturday, 31 January 2009 4:15 a.m.
>To: cytometry at lists.purdue.edu
>Subject: [Cytometry] Help with macrophage detachment
>
>Thank you in advance for your help with this issue.
>
>I am trying to detach human monocyte derived macrophages from tissue 
>culture dishes. I have tried Sigma's Cell Detachment Media and 
>Genlantis' Detachin and have considerable cell death and up to 20% of 
>the cells still remain on the plate. I am hesitant to use trypsin/EDTA 
>as I need to do surface staining of the cells for flow cytometry. I 
>have also tried mechanical removal (gentle scraping) but that also 
>caused a high percentage of dead cells. Thanks
>
>Doris Wiener
>University of South Florida
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