macrophage contamination in microglial culture - (Nov/14/2012 )

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!

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!

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.