Hello All,
I have been noticing that my cells haven't had the greatest viability. Typically when I passage my cells I see upper 90% in terms of viability and only see a drop in viability 3 days after I have reached terminal volume. However, recently I have seen lower viability when I passage around upper/mid 80% and then seemingly at random my cells viability drops into low 60% with very little cell density ~1e6. My first thought is contamination so I do the smell test and don't smell any sense of yeast. I check under the microscope at 4X, 10X, 20X and I don't see anything out of ordinary also checking with GFP lens and see that the cells that are clumped together are still showing green color. I check the images that are taken by the vi-cell and don't see anything out of ordinary. I have also been checking the culture visually and don't see anything that catches my eye. What else can I do to figure out what is happening with my cells?
Hi Bianka Maro
Can you share some photos of your cultivated cells, the composition of your medium and cell type?
Mohamed Khashan
I don't have any photos currently since I have already harvested the cultures,but the media is 293 freestyle media and the cells are 293F
Hi Bianaka
So
Viability is dependent on several factors
What is the cell under culture
- recommend not performing the smell test , that may cause contamination
- Do you see contamination in culture?
- Are you providing the essential requirements, in terms of medium with its required additives and change once in 2-3 days with appropriate volume .. 5 ml minimum for T25, 15 ml for T75 ?
- how do you estimate viability?
With countess?... Cross check that by manual method
- what is the passage number?
- using non expired reagents?
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