irradiated 3T3 cells as feeder cells - (Feb/24/2008 )
I am now culturing irradiated 3T3 cells and use them as feeder cells for primary lymphoma cells. I found that if I leave the feeder cells in culture for some time (without the lymphoma cells), say, about 1 week, the morphology will start changing and finally become ugly. I also found there are tiny particles accumulating around the cells, looking like the feeder cells are spilling out something.
My question is, if the morphology starts to change, can I still use the cells for feeder layer?
Does anybody know what the change means and what causes it?
I don't know lymphoma cell culture, but for ES cell culture, the feeder cells should be best used the next day after plating.
I usually plate 5.25x10E6 cells to get good coverage of a 10-cm plate. I found if I start using the feeder cells the next day, the feeder layer is very easy to be broken, even just by adding media dropwise. So I usually wait for at least 2 days to use them and by then the feeder layer becomes very firm.