My cloned cells are not supporting growth using the same media. It was kept in cryopreserved state in cell banks. Can any one help me to get an idea like how its possible? pH is maintained thus lactate accumulation condition has been ruled out.
Reviving cells that have been cryopreserved has to be done by carefully following the revival protocol. If the frozen cells are thawed in a 37oC water bath it is important that they do not reach 37oC and often the cryomedia contains DMSO which usually needs to be removed. What protocol are you following? Also, when the cells were initially frozen, did someone check a vial a week later for viability to make sure they were frozen properly? Improperly frozen cells will have low viability.
Dear Gary,
Thank you for the guidance.
You are right. Cells were not cryopreserved nicely by previous employee.
For removal of DMSO, we are centrifuging and and again re-suspending in new fresh media. And also taken care of Revival protocol as per your direction only has been followed.
Thank you for figuring out and helping me!
Regards,
Santosh Kumar
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