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