Enlarging Cells - (Aug/27/2007 )

greetings

im working with a culture of MDBK cells using DMEM/Hams F12 1:1 medium with 5% fetal bovine serum (non heat inactivated) at cosy 41°C. they are doing fine and usually reach confluency every 3 or 4 days. problem is that after reaching confluency the cells are very small and tightly clustered.

for my work i would rather need fewer and bigger cells with larger surfaces. do you have any ideas how this could be accomplished?

i was told that i should try serum starvation. by keeping the cells in G1/G0 phase and thus inhibiting mitosis the cells are supposed to grow larger than usual.

and someone else told me i should use a hypotonic medium and inflate the cells with osmosis!!

so do you have any tips? and would you agree or advise against the methods i mentioned here?

thanks a lot

what experiment do you hope to do on them??

I usually use my cells at approx 90% confluent for most experiments, as they are then still growing and not squished up by the cells around them (but I'm using a diff cell line to you)

im using them as host cells for infection with several protozoal parasites. thats why i dont want them to be cramped and small. the parasites should have enough space within the cells to multiply and develop.

Ok, then I would infect your cells with your parasite when they are around 80-90% confluent. Do your cells need to be still growing when you have infected them?? If not, then try reducing the serum concentration in your media down to 2%, this should be enough to keep them alive but stop them from dividing.

Hope that helps