[Cytometry] Frozen PBMCs for TBNK

Divya Bhatia divyabhatia2 at gmail.com
Mon Apr 1 01:10:18 EDT 2013


Hi,
I guess TBNK stands for T-cells, B-cells and NK cells, I did staining for B
cells (CD45+, CD19+), Tc (CD8+) Th (CD4+), and also cultured after
stimulation with PMA, Ionomycin and monensin for staining for Th1
(CD3+CD8-,IFN-gamma+), Th2 (CD3+CD8-, IL-4+,IL-13+) and for Th17
(CD3+CD8-,IL-17+,RORgt+) cells even after one month on cryopreserved cell
(stored at -80 degree). I found it would be better to stain within 15 days
after cryopreservation if you want to look for intracellular cytokines and
transcription factors. You ll get pretty good expression of surface markers
even after one month on cryopreserved cells.

Cheers
Divya


On Mon, Apr 1, 2013 at 7:07 AM, Sundararajan Jayaraman <anue2468 at uic.edu>wrote:

> It would be helpful if you explain what is TBNK. Also, how was the PBMC
> frozen, stored and how long? What is the yield of live cells after thawing
> in your hands?
>
> Thanks.
>
> --
> S. Jayaraman, Ph.D.
>
>
> On Tue, March 26, 2013 10:27 am, Lukas, Susan wrote:
> > Hi all,
> > Has anyone tried running TBNK on frozen PBMC's?  If so any pit falls?
> >
> > Susan
> >
> >
> > Susan Lukas, MPH
> > Sr. Biological Scientist
> > University of South Florida
> > Children's Research Institute
> > 601 4th Street, South CRI 4008
> > Saint Petersburg, FL  33701
> > 727.553.1259
> > 727.553.1295 (fax)
> > slukas at health.usf.edu<mailto:slukas at health.usf.edu>
> >
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-- 
*Thanks & Regards :
Divya Bhatia
Ph.D Student*
*Department of Pediatrics
All India Institute of Medical Sciences
New Delhi, India*
*Tel. 011-26546518*
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