MCF-7's Not Growing - (Oct/03/2005 )

I'm trying to grow MCF-7's using DMEM supplemented with glutamine, pen/strep and FCS. For a 150mm dish, I'm using 400uls cell suspension and 24.4mls medium. This has given a confluent plate before, but now all of a sudden the cell density isn't large enough to allow the cells to grow. The stock's also seem fine so I'm baffled as to what the explanation could be.

Any suggestions?

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Mycoplasma?

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did you try seeding at higher density? i.e. decrease your medium volume.

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Hi....
I use MCF-7 cell in my experiment same as you. For me, I using MEM+ 1% nonesssential amino acid + 10 % FBS(FCS).I think to solve your problem by increasing % of FBS up to 20-30% . Moreover, Are you sure that your cell is healthy when you put it up from liq N2 or -70 c . Maybe your cell is mortalized when you keep it in -70 C more than 3 months. and anouther problem when you thaw the cell, you should be done it not slowly!!!!!!!!!!!!!!!! because DMSO in freezing medium is toxic for cell in room temperature.

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I'm using MCF-7's as well. My media is RPMI with 10% FCS + 10ml Na/Pyr and 10ml pen/strep.

I had a problem reciently where I brought the MCF-7's up from frozen and they just sat on the plate and grew slowly. I changed the medium etc but nothing happened and they appered to stop growing completly for two weeks, never reaching confluance. So I tried sub cultureing them anyway. and they too off. It could just be they don't like the surface they are on.

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If you are not doing any hormone related work....go ahead and add 10 nM Estradiol to your plates. You will get a lot more cells that way.

Good luck
p..

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