Hi,
I have noticed that my HEK293 cells become round shaped when I grow them on poly-L-lysine coated coverslips. They are not dead, I seeded the same amount of cells from the same tube on coated and uncoated coverslips and checked the cells after a few h - on uncoated coverslips they have their characteristic shape (but not a lot of them were attached), on the coated coverslips - they were round (but attached).
Is this possible that using PLP change morphology of my HEK293? Do you know any other reagents I could use to coat coverslips for HEK293 without this morphology-changes effect? Which reagent do you use for HEK293, which is the best one?
Hi the same happened to me, so I am using HeLa cells, also glass wells are better for microscopy.
HEK 293 cells can grow in a normal polysterene tissue culture flasks or plates .these are supplied by various cell product supplying companies .No need for extra coating of its surface.See that you are adding all the necessary components required for the growth of the cells.
A lot of factors influence cell morphology, such as media components and growth surface. As mentioned by Dr. Reddy, HEK293 cells are often grown on flasks without any coatings; I have done this many times myself. I doubt the poly-L-lysine is having any negative impact on the cell viability; they are just appearing different as a result of the coating. You can either stick with the coating and see how your visual/assay results are impacted, or accept the loss of cells on an uncoated slide and proceed from there. I am not familiar with how HEK293 cells appear on other coatings; you can try a few different types and see which one comes closest to giving you the "expected" appearance.
I have the same problem. According to my PI, I should have washed the wells after coating to remove residual unbound poly-lysine. He thinks possibly unbound content is sticking to the cells and preventing them from attaching to the actual surface.
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