RAW mouse macrophages floating in culture - (Mar/22/2008 )

Hi!

I have been doing cell culture of RAW mouse macrophages for 2 years now and thought that I can figure out many of the problems. But probably not this one!! Whenever I split my cells these days, I see that there are many dead cells and they are floating, like say approx 20%. I tested the culture without antibiotics and there is no contamination coming up. I am scraping the cells but I am sure I am not doing it too hard to kill the cells. Any ideas why it could be??


Thanks

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Dear repeatcell,

We grow our RAW cells in stirrer bottles....why?

No need to scrape or trypsinise them for experiments or passaging.....always 99% viability.

The stirrer bottles are re-usable....therefore long term you save loads of money.

You can grow HUGE numbers of cells in a small area.....therefore saving incubator space.

By not trypsinising or scraping, receptor expression experiments can easily be done.

TIME SAVED IN PASSAGING....pour off cells, pour in fresh media.

Hope you find this useful.

Kindest regards

Rhombus

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are you sure that the same receptors are expressed on the surface when a macrophage is not adhering, may be it loses some of the receptors/express them at a low density when grown under stirring conditions??

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