Hello
Currently I'm working on a permeability study using Caco-2 cells.
I have been working on cells for nine months to study their behavior and increase the number of the passage, but in some flasks and around the month of (August) I started noticing a very fast growth of cells for a month and then at the beginning of the month (September) their growth became slow until now more severely.
On the high magnification (40X) I noticed some sticky black dots that I don't know exactly what they are (I assumed it was the cells debris) while there was no turbidity (only a slight turbidity) or a change in the color of the media.
Fearing that these spots were contamination of the cells, I put gentamicin, which helped me at first, but eventually the sticky black dots came back and the cells came back slow and weak, and I stopped applying it.
Have you any ideas or suggestions ?
Thanks in advance.
I attached images for the cells:
1)In the beginning (p=26)
2)In the Rapid growth period
3) Before / After applying Gentamicin
5)And an image for cells under Confocal microscopy with DAPI stain
Check on this paper
https://www.vanderbilt.edu/viibre/CellCultureBasicsEU.pdf
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