[Cytometry] Cell cycle of activated human PBMC

Gary Warnes g.warnes at qmul.ac.uk
Mon May 23 11:13:40 EDT 2011


Dear Marjan,
	In my experience naive human T-cells show little detectable cell division, they must zip through the cell cycle very quickly, in this scenario even at 3 days there is little detectable cell division by this method, so I am not surprised by your result. I guess CFSE proliferation gives you more usable results given that the S phase & G2m percentages are at any one time point very low.

Hope this suggestion helps

Gary 

Dr Gary Warnes
Flow Cytometry Manager 
Flow Cytometry Core Facility
Blizard Institute of Cell & Molecular Science
Barts and The London School of Medicine and Dentistry
London University
4 Newark Street 
London
E1 2AT
Tel No.0207-882-2402
Fax No. 0207-882-2180


-----Original Message-----
From: cytometry-bounces at lists.purdue.edu [mailto:cytometry-bounces at lists.purdue.edu] On Behalf Of Marjan Gharagozloo
Sent: 22 May 2011 09:53
To: cytometry at lists.purdue.edu
Subject: [Cytometry] Cell cycle of activated human PBMC

Dear All,
I'm working with human PBMC, activated by 2µg/ml CD3/CD28 mAb. I've
performed cell cycle analysis after 24h activation on these cells and I've
got strange histograms! didn't see S phase! I have no idea about the
populations in FSC/SSC dot plot; I can see 3 populations, where I should
gate cells? What is the problem with this test?
Please see attached file. Any suggestion and comment would be greatly
appreciated.

Looking forward your kind reply

Best regards:
Marjan Gharagozloo (Ph.D)
Assistant professor
Department of Immunology
School of Medicine
Isfahan University of Medical Sciences
Isfahan, Iran























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