Hello my hopeful rescuers!
I'm working with RAW264.7 cells in RPMI1640 media. I seeded them in a 96 well plate with different concentrations starting with 1*10^4 cells/100 µl to 5*10^4 cells/100 µl. As you might have guesses the volume per well was 100 µl. Most of the papers I found on this subject incubated at 37 °C and 5% CO2 for 24 h before they started with the assay. My problem now is that even with the highest concentrations of cells I cannot reach confluency even after an incubation of 48 hours.
My question or let's call it my cry for help concerns your experiences with culturing RAW264.7 cells in micro titer plates. What was the highest confluency you could reach? How long did you incubate the plates? What was the initial concentration of cells you used?
Hope someone out there can help me
yours desperate Jules
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