I have revived Jurkat E6.1 cell lines by removing the freezing media (10% DMSO in FBS) by complete medium. However in general practice the cell debris will stop to recurring after one passage. But they are still appearing in the medium. What would be the cause and how to overcome it?
The debris often happens by de-freezing the cells and usually it stops after one passage. I guess you are using appropriate medium but you should check viability of the cells and to calculate the number of living cells. It is one the way how you will be able to check to percent of living/dead cells. By time, the percentage of the dead cells should be lower. Try to passage them few times.
Have you determined post-thaw viability by trypan blue exclusion test immediately after thawing of cells? It's usually recommend to be >70% living cells.
Could you give more detail about your freezing? Maybe you should optimize the freezing procedure to lower amount of dead cells and debris after thawing.
Additionally, you could add an extra medium exchange before subcultivation in order to remove cell debris. For adherent cells it is recommend to have a 24 hour plating peroid before changing cell culture medium.
If the problem persists after several passages, I guess it is most likely not a problem of the freezing or thawing procedure. What medium are you using for your Jurkat and at what density do you keep them? In my cultures I noticed that jurkats do not like to be diluted too much. Also FCS can heavily affect your Jurkat cell culture. Maybe it would be worth to test different batches of FCS for your cell culture.
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