Re: Storage temperature for freshly separated PBMC

From: Mathi (mathix@meyerpharm.com)
Date: Thu Mar 02 2000 - 19:17:11 EST


Hello Wim

Ficoll is a non specific activator. If you store at 37c at 5% CO2 your
cells will see activation the next morning. I would store the whole
blood. Or buffy coat the cells and store them over night and ficol next
morning if the procedure demands ficolling. I work with Apheresis
material. i store them overnight with
plasma, no problem so far. Apart from all these my experince says auto
activation in over night incubation is also very much real.

Thanks

mathi

Wim Jennes wrote:

> In my intracellular staining protocol, PMA/ionomycin stimulation
> is usually done the day after blood collection and PBMC ficoll
> separation. Does anyone know what is the best overnight storage
> temperature for PBMC? I did 37 °C, 5 % CO2 and also closed tubes at
> 4°C, both in RPMI supplemented with BCS. For certain cytokines these
> 2 conditions really give different results!
> Thank you for helping,
> Best regards,
>
> Wim Jennes
>
> ---------------------------------------------------
> Wim Jennes
> Institute of Tropical Medicine
> Department of Microbiology
> Laboratory of Immunology
> Nationalestraat 155
> B-2000 Antwerpen - Belgium
> Tel: + 32 (0)3 247 62 27
> Fax: + 32 (0)3 247 62 31
> e-mail: wjennes@itg.be
> ---------------------------------------------------



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