Hi, I have to culture AML12 cells in a 6-well plate for 24hours followed by treatment for another 24 hours (total experimental duration=48 hours). The cells will be lysed at the end of the experiment and used for metabolomics (need >=1 million cells/well at the end).
Earlier, I seeded 0.5 million cells per well with 2mL media in each well. However, the cells were not 50% confluent after 24 hours. I wish to know the cell seeding density or cell number of AML12 cells per well in a 6-well plate so that they are about 60% confluent after 24 hours.
Bhaskar Ganguly 4 million cells per well of a 6-well plate? Won't it exceed the well capacity?
Please work out on population double time and viability for the mentioned cell line AML-12 and compatibility of culture media (for growth and adherence). Seed a known number of viable cells < 1 million cells (depending on doubling time) on Day 0, to treat million cells next day (after 24 hrs).