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macrophage contamination in microglial culture

macrophages macrophages microglia microglia astrocytes astrocytes M-CSF M-CSF GM-CSF GM-CSF

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#1 mken

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Posted 14 November 2012 - 08:54 AM

I am prepping glial cells from neonatal murine brains, to split them and investigate astrocytes and microglia separately.

Here's what bothers me (due to my nature of questioning everything):

The way we prepare the cells in my lab, we culture them from whole brains, and there is usually some blood contamination at the start of the culture.
From what I know the same growth factors that enhance the growth of microglia, are also macrophage growth factors (we use M-CSF and GM-CSF). How can I be sure that what I have in my culture is actually a microglial cell and not a macrophage? I asked a fellow from my lab about this and they claimed that macrophages would not attach to the bottom of the culture flask, whereas microglia do, so I can be sure that what I have are micros and not macros (the macros would be discarded in the supernatant upon medium change). However from what I read about culturing macrophages, it seems to me that cultured macrophages also attach to the bottom of the flask.

Anybody has any idea about this?

Thanks a million!

#2 mken

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Posted 14 November 2012 - 08:55 AM

I am prepping glial cells from neonatal murine brains, to split them and investigate astrocytes and microglia separately.

Here's what bothers me (due to my nature of questioning everything):

The way we prepare the cells in my lab, we culture them from whole brains, and there is usually some blood contamination at the start of the culture.
From what I know the same growth factors that enhance the growth of microglia, are also macrophage growth factors (we use M-CSF and GM-CSF). How can I be sure that what I have in my culture is actually a microglial cell and not a macrophage? I asked a fellow from my lab about this and they claimed that macrophages would not attach to the bottom of the culture flask, whereas microglia do, so I can be sure that what I have are micros and not macros (the macros would be discarded in the supernatant upon medium change). However from what I read about culturing macrophages, it seems to me that cultured macrophages also attach to the bottom of the flask.

Anybody has any idea about this?

Thanks a million!

#3 bob1

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Posted 14 November 2012 - 01:09 PM

It looks like you hit the post button twice - so merged threads...

Macrophages do attach to the flask. I think you should be able to distinguish them (quite easily, having done a google image search) based on morphology of the cells.





Also tagged with one or more of these keywords: macrophages, macrophages, microglia, microglia, astrocytes, astrocytes, M-CSF, M-CSF, GM-CSF, GM-CSF

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