[Cytometry] Help with macrophage detachment

Thure Adler thureroman at gmx.de
Thu Feb 5 09:00:00 EST 2009


hello, 
I once detached macrophages with the help of the product 'Accutase' which is considered to be 'softer' than Trypsin/EDTA.
See this protocol also in: 
Heat shock protein 60 elicits abnormal response in macrophages of diabetes-prone non-obese diabetic mice.
Adler T, Akiyama H, Herder C, Kolb H, Burkart V.
Biochem Biophys Res Commun. 2002 Jun 14;294(3):592-6.

-------- Original-Nachricht --------
> Datum: Wed, 4 Feb 2009 13:30:34 +1300
> Von: "Parlane, Natalie" <natalie.parlane at agresearch.co.nz>
> An: "\'Wiener, Doris\'" <dwiener at health.usf.edu>, "cytometry at lists.purdue.edu" <cytometry at lists.purdue.edu>
> Betreff: 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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-- 
Dr. med. vet. Thure Adler
Immunology Screen/GMC at the GSF
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