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THP-1 cell surface markers for Flow Cytometry - Need a robust marker for undifferentiated THP-1 cells (Sep/24/2009 )

I am working with THP-1 cells (ATCC TIB 202), a human monocytic cell line. The majority of people that use this cell line are interested in the differentiated macrophage phenotype, which can be achieved by stimulating THP-1 cells with PMA or Vitamin D.

I mainly use these cells in the "undifferentiated form" without any type of stimulation. I am planning on doing some cell surface expression of various molecules using flow cytometry. I have several different fluorophores and need to run a positive control to compensate for the spectral overlap between the different fluorophores.

Unfortunately, the majority of the literature characterizes cell surface expression of PMA/Vitamin D differentiated THP-1 cells which can tremendously affect the cell surface expression. One of the common markers used is CD14, which is expressed in extremely low levels in undifferentiated THP-1 cells.

I was wondering whether anyone could recommend some markers for undifferentiated THP-1 cells.

Thanks a lot.

I am new to flow cytometry.

-BigKahunaBurger-

Hi,

Sorry but I don't have any helpful info for you, i have just started working in this area. I was just wondering whether you had found any useful markers for the undifferentiated form?

Thanks

BigKahunaBurger on Sep 25 2009, 03:04 AM said:

I am working with THP-1 cells (ATCC TIB 202), a human monocytic cell line. The majority of people that use this cell line are interested in the differentiated macrophage phenotype, which can be achieved by stimulating THP-1 cells with PMA or Vitamin D.

I mainly use these cells in the "undifferentiated form" without any type of stimulation. I am planning on doing some cell surface expression of various molecules using flow cytometry. I have several different fluorophores and need to run a positive control to compensate for the spectral overlap between the different fluorophores.

Unfortunately, the majority of the literature characterizes cell surface expression of PMA/Vitamin D differentiated THP-1 cells which can tremendously affect the cell surface expression. One of the common markers used is CD14, which is expressed in extremely low levels in undifferentiated THP-1 cells.

I was wondering whether anyone could recommend some markers for undifferentiated THP-1 cells.

Thanks a lot.

I am new to flow cytometry.

-panda-