[Cytometry] about CD14-negative mature monocytes

Nuno Oliveira cravista at gmail.com
Fri Feb 21 19:06:32 EST 2014


Hi

I sent you some documents to read. You have a lot of answers there. Some of
them are in other languages like spanish, portuguese or dutch, but you can
see tables or histograms for exemple, which are easy to understand, even in
foreign languages. If you need some translations you just go to dictionary
"language of the document - your language" (in this case english) and you
can find good answers for sure. Let me know if those documents are helpful.

Best,

Nuno Oliveira, IPOCFG - Flow Cytometry Division



2014-02-21 23:44 GMT+00:00 Tuluc, Florin <TULUC at email.chop.edu>:

>  Hi Nuno,
>
>
>
> Thank you for your message - it is helpful. I am familiar with the papers
> by Ziegler-Heitbrock and others (you probably know the review article
> attached). They describe CD14dim and CD14bright subpopulations of monocytes
> but I do not recall seeing any paper that looks at CD14-dead-negative
> monocytes.
>
>
>
> I do see in my samples cells that are CD14-negative and CD16-bright (but
> they are back-gated in the lymphocyte gate on the FSC/SSC plot). Why do you
> think one can call those cells monocytes based on the expression of CD300e,
> CD64, and/or CD36? I am interested to learn more on this topic and if you
> are aware of any paper describing CD14-negative mature monocytes please let
> me know.
>
>
>
> Best,
>
> Florin
>
>
>
>
>
> *From:* Nuno Oliveira [mailto:cravista at gmail.com]
> *Sent:* Friday, February 21, 2014 5:06 PM
> *To:* Tuluc, Florin; cytometry at lists.purdue.edu
> *Subject:* Re: about CD14-negative mature monocytes
>
>
>
> Hi Nuno,
>
> I saw your message on the Cytometry list. I wanted to ask if you know what
> percentage of mature, circulating monocytes are CD14-negative and how can
> one prove that those CD14-negative cells are indeed monocytes. If you know
> of a paper that describes these cells please let me know.
>
> Thank you,
> Florin
>
>
>
>
>
> >>>>>>>>>> Hi,
>
>
>
>
>
> I will answer below.
>
>
>
> As far as I know, the distribution of monocytes in the peripherical blood
> is this:
>
> - 80 % are "classical" monocytes with CD14+ CD16-;
>
> - 10 % are "non-classical" monocytes with CD14- CD16+;
>
> - 10 % are, let´s call "intermediate" monocytes with CD14+ CD16+ >>> this
> population can be divided in two subpopulations, depending on the
> expression of HLA-DR: one has high expression and the other one not so high;
>
>
>
> All (or almost all) of the monocytes are IREM2 positive (CD300e) and CD64
> positive as well. I think that all of them are CD36 positive as well...
> Dendritic CD16+ cells also express IREM2.
>
>
>
> So, if you want to study PB monocytes and you just have 4 or 5 colours,
> just put CD14, CD16, HLA-DR and CD45.
>
>
>
> I hope it helps.
>
>
>
>
>
> Greetings,
>
>
>
> Nuno Oliveira, IPOCFG - Flow Cytometry Division
>
>
>
>
>
>
> 2014-02-21 14:59 GMT+00:00 Tuluc, Florin <TULUC at email.chop.edu>:
>
> Hi Nuno,
>
> I saw your message on the Cytometry list. I wanted to ask if you know what
> percentage of mature, circulating monocytes are CD14-negative and how can
> one prove that those CD14-negative cells are indeed monocytes. If you know
> of a paper that describes these cells please let me know.
>
> Thank you,
> Florin
>
>
> Florin Tuluc
> Flow Cytometry Core Lab
> The Children's Hospital of Philadelphia Research Institute
> Office: 267-426-5350
>
>
>
>
> -----Original Message-----
> From: cytometry-bounces at lists.purdue.edu [mailto:
> cytometry-bounces at lists.purdue.edu] On Behalf Of Remy HALLET
> Sent: Thursday, February 20, 2014 3:00 AM
> To: Vyzasatya Ravi - Lonza Walkersville
> Cc: cytometry at lists.purdue.edu
> Subject: Re: [Cytometry] CD14 positive cell line
>
> Hello there,
>
> I've been looking for a CD14+ cell line too.
>
> In my hands U937 and ThP1 were negatives. I found Hodgkin Lymphoma KM-H2
> cells slightly positive (medFI x3 compared to isotype control, very weak
> compared to freshly isolated monocytes), as indicated on DSMZ data sheet.
>
> I didn't try Mono Mac 6.
>
> Hope this can help.
>
>
>
> Rémy Hallet
>
>
> Chargé de Travaux Scientifiques
>
>
> 03 68 33 28 28
>
>
>         E-mail : hallet @transgene.fr
>
>
>
>
>
>
>
>
>
> Transgene S.A.
> Boulevard Gonthier d'Andernach
> Parc d'innovation
> CS80166
> 67405 Illkirch-Graffenstaden Cedex - FRANCE Tel +33 (0)3 88 27 91 00 Fax +33
> (0)3 88 22 58 07
>
> http://www.transgene.fr
>
>         _________________________________________________
>         Pensez à l'environnement avant d'imprimer ce message.
>
> ----- Mail original -----
>
> De: "Vyzasatya Ravi - Lonza Walkersville" <ravi.vyzasatya at lonza.com>
> À: "Ina Laura Pieper" <inalaurapieper at gmail.com>
> Cc: cytometry at lists.purdue.edu
> Envoyé: Mercredi 19 Février 2014 17:59:32
> Objet: Re: [Cytometry] CD14 positive cell line
>
> Exactly..
> Tried THP-1. It is negative for CD14.
> As Tracey Duff suggested Mono Mac 6 may be my best bet.
>
> RV
>
> From: Ina Laura Pieper [mailto:inalaurapieper at gmail.com]
> Sent: Wednesday, February 19, 2014 4:52 AM
> To: Nuno Oliveira
> Cc: Vyzasatya Ravi - Walkersville; cytometry at lists.purdue.edu
> Subject: Re: [Cytometry] CD14 positive cell line
>
> Do mature monocytes proliferate in culture though? I think Ravi may be
> looking for a cell line that can be maintained and harvested whenever a
> biological CD14+ control is needed for flow?
> I've got daily access to blood samples so I use the CD14+ monocytes in
> whole blood as a control, but I would be very interested in learning more
> about CD14+ cell lines too.
>
> Best wishes
> Ina Laura
>
> On 18 February 2014 20:56, Nuno Oliveira <cravista at gmail.com<mailto:
> cravista at gmail.com>> wrote:
> Hi
>
> If I understood your question, the answer is mature monocytes. Almost all
> of them express CD14.
>
> Greetings,
>
> Nuno Oliveira, IPOCFG - Flow Cytometry Division
>
>
> 2014-02-18 13:56 GMT+00:00 Vyzasatya Ravi - Lonza Walkersville <
> ravi.vyzasatya at lonza.com<mailto:ravi.vyzasatya at lonza.com>>:
>
> > Dear Flowers,
> >
> > Do you know of any cell line(s) that constitutively express CD14? I am
> > looking for a CD14 cell line that I can use as positive control in a
> > flow assay.
> >
> > Thanks in advance.
> > Ravi
> >
> > Ravi Vyzasatya
> > Scientist
> > Cell Therapy Development Services
> > Lonza Walkersville, Inc.
> > 8830 Biggs Ford Road
> > US - 21793-0127 Walkersville
> > Tel. : +1 301 378 1152<tel:%2B1%20301%20378%201152<%2B1%20301%20378%201152>
> >
> >
> >
> >
> >
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