What is the recommended seeding density for H9C2 cells (specifically in a plastic tc t25 flask)? We have been seeding at the recommended 10x10^4/cm^2 (250,000 in t25) and they are reaching ~70% at 48 h instead of 72 h. The majority of our cells seem to be attaching. Can we seed at lower amounts without changing the cells? Additionally, how many cells would be in a confluent t25? We are using the media and trypsin specified by ATCC.
Hi,
I have been culturing h9c2 for a couple of months in T25 flask and at 70% usually it reached 420,000 cells per well. And the population doubling time for H9C2 is 2.3 days if it grows well. So if I want it to achieve confluency at different day, I seed the cells according to split ratio as follows:
1: 2 (210,000 cells) = 2 days
1:3 (140,000 cells) = 3 days
1: 4 (105,00 cells) = 4 days
Hope this helps
Thank you
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