I am cultivating right now MEG-01 cells in the following medium
RPMI1680 from Sigma Aldrich
5 mL Pen-Strep-l-Glutamine Sigma Aldrich
50 mL FBS from GIBCo
While the last half year the cells were happy and grew fast and well, I have a lot of trouble now. It all started with people changing the medium supplements without telling and out of sudden my cells died. So now, 2 batches later, I have tried to find the former formualtion of the medium (which is the one above) and it looks better than before. The cells don’t die immediately. Instead it took them last time 1 week until I realized that many cells died and then all were killed. Since I was not sure if bacteria were the reason I prepared new medium and I defroze a new batch, believing that it will be better this time. Lucky me, the viability of the new defrozen cells is only about 50 % (??? vials from the same stock had 80-90 % viability before and I had no trouble in growing them!) and they start dieing again. You can see the dead material in the flask.
Now I am assuming that this time the cells will die again as the last times. I will make some tests now if there are any bacteria in there (though the medium was also used for other cells which were not infected!).
But I have only one batch of this cell line left and I was wondering if there is anybody who has handled this cell -line and can give me some advise. Or if there are any possible reasons for the dead that I don’t see?
Please let me know what I can do…I am feeling totally frustrated.
I think I have the same problem with MEG 01. I defrost them 3 days ago and they are with many fragments suspended in media with some attached cells. I did not notice an obvious in growth until today. This is the first time I work with MEG 01. How could I differentiate If my cells are dying or this is normal.