Hello All, I am currently using 3T3-A31 (ATCC) cells for an assay. The assay is about 12 days and I am continuously facing an issue with the cells in the assay plate, as they are getting peeled off from the plates or forming clusters in the assay plate. The cells are healthy and good enough before addition of differentiation media and after addition of it, we are observing the same pattern as shown in the pictures. Could you please let me know what might be the issue? Thank you.
Hi Priyanka Ghantasala . You often see this separation of cell masses in overconfluent cultures where cell-cell contacts are stronger than cell-substratum contacts.
Use less dense cultures for your experiments.
Steingrimur Stefansson brings up a good point in terms of confluency. Additionally, it would be beneficial if you can provide further details- what you are trying to differentiate them into. Any information on the culture media composition, the substrate material, and coating used would also aid in providing a better answer.
Thank you for your response. But I have tried cells from 15000 cells to 30000cells per well. Even though, I have observed the same pattern. The differentiation media contains DMEM+ 0.25 µM Dexamethasone + 0.5mM IBMX + 1 µg/mL Insulin.
- Badges
- Science topic
- Similar topics
- Thesis Research