Dear researchers,
I am currently culturing THP-1 cells in a T75 flask. I would like to inquire whether it is possible to achieve a confluence of 100% with 32 million THP-1 cells in 20 ml of medium, considering that THP-1 cells do not adhere to the flask.
Thank you for your responses.
Hind
Hello Hind Sebbah
No. The count of 32 million cells in 20ml of media would be a bit too much, and I would term it as over confluent and not 100% confluent. Cells would begin to clump together and float on top of the media. You would also notice the media changing color slightly and appearing more turbid. If the media turns acidic by looking at its color, it will indicate that there are too many cells in culture.
Please note that healthy growing suspension cells should be round and bright, with minimal cell debris. I feel that you should have 8-10 x 10^6 cells in 20 ml of media. THP-1 cells are very sensitive to cell density. The culture conditions of THP-1 such as concentration and time is extremely important if you wish to use these cells for further processing.
You may want to refer to the paper attached below for more information.
Article Effect of Culture Conditions on the Phenotype of THP-1 Monoc...
Best.
Hello Malcolm Nobre ,
Thank you for your response! I now have a better understanding as I believe my THP-1 cells were overconfluent. The media remains unchanged. Ah, I see, 8-10 x 10^6 cells in 20 ml of media.
I have another question: when placing the cryovial tubes in Mr. Frost for freezing, should I leave them slightly open or fully sealed?
Best regards,
Hind Sebbah
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