[Cytometry] about CD14-negative mature monocytes

Nuno Oliveira cravista at gmail.com
Fri Feb 21 17:06:03 EST 2014


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
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> 67405 Illkirch-Graffenstaden Cedex - FRANCE Tel +33 (0)3 88 27 91 00 Fax +33
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>         _________________________________________________
>         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>
> >
> >
> >
> >
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