hi again! I had never worked with Caco-2 cells before and just recently I thaw some. When I look at them I see something that I really don't know what it is. I know these cells secrete a lot of mucus and that it is observed at the confluence, but I don't know if the little spots in the culture -as observed in the picture- are normal. So, is it contamination?
Hi Melissa Tirado
It seems that you may be concerned about the particle-like substances in your cell culture. However, based on the timing of your image capture (it seems right after thawing the cells), it is difficult to determine whether these particles are simply cell debris (which is often present after thawing cells, depending on the quality of the prepared stock) or if they indicate contamination.
It's worth noting that Caco-2 cells typically grow in clusters (use 20% FBS) and take some time to proliferate after thawing. If you continue to observe these particles after the cells have had a chance to grow and the media has been changed, there may be cause for concern about contamination.
Best wishes,
Tanoy
Hello Melissa Tirado,
The image shows poor cell viability. Have you checked the percent viability after thawing? If you have fewer cells in culture, the chances of revival are less.
Tanoy is right. Usually cell debris are noticed after thawing. So, the tiny spots that you observe are most likely to be cell debris which may be removed by changing the media with fresh media. If you have a doubt whether the culture is contaminated, you could always streak a few microlitres of the culture media on agar plate and incubate the plate at 37 degree C overnight. If the culture is contaminated, you will observe bacterial colonies growing on the plate the next day.
I feel you should check your freezing protocol and the storage conditions. These may not be properly followed due to which there is a loss of cell viability.
Best.
Hi Melissa Tirado
First, Caco-2 cells do not secrete a lot of mucus since they do not have goblet cells.
However, according to the image and after cell thawing it may observed these dots, which are debris that should disappear within the first week after thawing.
If it remains for a longer period, there is a greater possibility of cell contamination, which in turn increases cell death.
Note: Cells need to be present in a large number because they need to be close to each other for their growth to increase.
Best regards,
Mohammad
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