I have seeded 50,000 NTERA cells per well in a 96-well plate in preparation for a scratch assay. I seeded the cells on the 11th of September. And even after 48 hours (13th of September), some of the cells have either withdrawn from the edges of the well or are simply not growing there. My questions are;
How did this happen? (Do the monolayers peel off the edges sometimes?)
Is there any way to avoid this in future scratch assays?
Would there be an issue if I scratch the monolayer while there are a few gaps in the edges?
For context I am using NTERA-2 cells (passage 12), 50,000 cells per well, seeded on a 96-well cell culture treated plate, flat bottom.
Hello Navodya,
First of all, it is always better to use a 6 or 12-well plate to perform a scratch assay. Then seed the number of cells as per their doubling time. Otherwise, they will get over-confluent and the edges will chip off (as in the figure).
Regards,
Nilanjana
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