I seeded 5e6 and 3e6 SW756 cells in two 15cm dishes with equal amounts of media on Thursday, anticipating one to be confluent Monday/Tuesday, and the other to be confluent sometime later, maybe Thursday. This is my first time testing with these, so it was just a rough guess.
However, on Monday, both were equal confluency - I could not have told a difference in plates if they weren't labelled. Is this normal? Should I give less media to the 3e6 plate to get the anticipated effect, or just plate less cells next time?
Just overall curious what factors play a role in cell growth, as the 3e6 plate grew at a much faster rate with the same conditions - aside from having a higher proportion of fresh media.
Hello Ethan,
As per the information provided by you, there could be two possiblities.
1. You may have wrongly counted the cells while plating, or
2. You may have wrongly concluded the confluency state of cells.
I would infer that 15cm dish with 5e6 seeding density may have become overconfluent on Monday because of which there is lack of space for cells to grow further as well as the media may have been depleted of nutrients.
On the other hand, the 15cm dish with 3e6 seeding density may have almost reached confluency with some space left for cells to grow besides the media still being fresh and nutrient- rich.
I would suggest you plate less cells this thursday (say 3e6 so that it reaches 80-90% confluency after 4 days i.e., Monday/Tuesday and 1.5e6 cells so that it reaches near confluency at one/two days later i.e., Wednesday/Thursday) with the SAME amount of media in both the 15cm dishes.
Best.
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