Hela cells are cultured with DMEM+10%FBS+antibiotics in our lab using nunc 25-TC bottle. However, cells look awful (as attachment). But when Hela cells was plated in nunc 24-well plate and cultured overnight, and change for fresh DMEM complete media like for 2~4 hrs, Hela cells looks better (smooth, as attachment). considering that the cells may be contaminated, the cells were culture without antibiotics for 2 passages, no contamination was observed and the cells look the same. Wondering if you have any similar experience.
On a fresh plate the cells are a little less spread so they are more smooth (lower amount of filopodia) and the light disperses a little better on the cell borders so they are a little bit more shiny. Tumour cells are usually much less attached than the normal cells but they still produce elements of extracellular matrix and when the produced proteins accumulate on the plate then tumor cells spread a little better. On a fresh plate they usually are just the slightest bit less attached.
Patrycja, thanks for your reply. The issue confused us for a long time. When the Hela was thaw from LN, they looked just fine, However, after 5~6 passages, they just looked like as attachment (a lot of filopodia and cell borders not smooth).
However, I still wonder the reasone that cell become smooth after media change is because of extracellular loss?
If You want to get the theoretical basis for this change it would be the best if You have looked through this three books, especially the one under the redaction by Rovensky. They nicely explain the changes in the cell shape due to the cell-ECM interactions (and not only this).
Rovensky - Adhesive Interactions in Normal and Transformed Cells, 2011
Mecham - The Extracellular Matrix - Overview, 2011
Gefen - Cellular and Biomolecular Mechanics and Mechanobiology, 2011
Patrycja, Thanks for your reply. However, the issue i face may be simply contamination, because several round white plaques were observed at the interface of media and side wall of culture bottle after the Hela was culture for 2 days in complete media without antibiotics. I dont know what is the contamination,
surprisingly, the media did not turn yellow.
Contamination and the resulting metabolites and shedded products can also influence cell attachement through, e.g., interactions with cell surface receptors inducing their aggregation (similarily like decorin induces aggregation of growth factor receptors). Also cell death induces deattachment.
And media turns yellow only if contamining organisms decrease the pH of the medium and it does not always happen.
It is good that You have now quite clear cut explanation for the changes. It is much worse if You have to analyze if the change is the result of the new source of plastic, media, FBS or different cell treatment (e.g., overtrypsynization).
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