I started a cell culture with a mix of suspended and adherent cells (presumably fibroblasts) on May 10th. After a week, I treated them for adherent cells only. On May 24th, most of the adherent cells appeared round and not the typical fibroblast shape, although some fibroblasts were still present. The medium looked clear and there were no signs of contamination.
You can see in the pictures, the one I took on 10th of May, and then the one on 24th of May.
Are these round cells healthy, or should they all be elongated fibroblasts?
Details for Researchers:
- Cell type: Fibroblasts
- Treatment change: May 17th (treated for adherent cells only)
- Observation: May 24th (mostly round cells with some fibroblasts)
- Medium: Clear, no turbidity
- No signs of contamination
It's very hard to see any cells, but I would say that all your fibroblasts are gone. Are the round cells attached?
Even in the first photo, they are very sparse and in my experience, cells like to be closer together in order to be happy. Was the plate at least 50% confluent when treated?
Dear Kerry S Vistisen thanks for your response. Yes, the round cells are attached. Furthermore, there are fibroblasts as well but overall the confluency is not close to 50%. I am following the proper protocol but my cells are not growing enough. It's been almost 2 months and the cells are still not 50% confluent.
Confluence is too low (top) to supply enough growing factors between them... Strange because it usually very easy to culture fibroblats - they are so invasive... Use a smaller petrish when seeding if you can't get enough cells (as 12 wells plate or less). I unfortunatly think cells are dead or will die (bottom). Do a trypan blue to confirm, as anyway, they will not survive. Good luck.
Dear Suzanne Cossette thank you so much for your answer. Yes, I think doing trypan blue will be a good way to confirm whether my cells are alive or dead.
Furthermore, I would like to ask that why my cells (fibroblasts) are not increasing in number ? I have noticed increase in round cells (adherent) but not fibroblasts.
Hi Bilal. What I and Suzanne were trying to convey is that you need to start your cells in a smaller dish so that they are closer together (more confluent) to begin with. Cells will not divide or will divide very slowly if they are not happy and happy cells have friends that are close to or touching each other. Once they are confluent in the smaller dish, then you can plate them in the larger dish in order to do your experiment.
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