[Cytometry] Help with macrophage detachment

Paul W. Price pwprice0701 at earthlink.net
Thu Feb 5 13:45:08 EST 2009


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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