[Cytometry] Help with macrophage detachment

Parlane, Natalie natalie.parlane at agresearch.co.nz
Tue Feb 3 19:30:34 EST 2009


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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