I have a serious problem. I've lost all the banks of BeWo line. I'm afraid to check other lines. After rebanking next day all cells are smashed. I note that we did a lot of experience using this line and particular banks were frozen at different times. We excluded individuals, the media, etc.We work with different cell lines over 20 years and the first time happened something like this.
I ask for suggestions .
Wasn't there any problem with the freezer? Freeze and thaw cycle could probably cause that.
Sorry - I can't help - I have no experience with these cells. Good luck!
Hi..when you said lost and smashed, is it that there is contamination or cells are physically damaged (under microscope?) or they simply are not growing....You will have to carefully use elimination process to zero in on actual culprit
Have you checked the freezer for possible errors or blackout? If the problems happen with all the cells in the bank maybe the problem is the criobank... levels of liquid nitrogen, temperature or power supply
Thank you all for your willingness to help.
We ruled out all the opportunities. Cells are in Dewar with liquid nitrogen, supplemented regularly. Were seeded in different media in order to exclude the media as the cause. Tests on mycoplasmas are negative. Usually we progresses according to procedure. Media have been added to the partially frozen cells, and incubated till attachment, then change media for fresh.
We tried also removed DMSO by centrifugation and cells were incubated in fresh medium. Effect was the same.
At this time, all JEG-3 banks have gone. One bank BeWo, I afraid to jinx looks good. What's surprising on those cells we conducted study in October and JEG-3 and BeWo were taken on exercises with the students as indestructible.
The causes we do not know and therefore purchase of the next line will give us nothing.
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