Hi,
We culture neural progenitor cells in T175 culture flasks coated with PLO-Laminin.
We have successfully grown these cells with our protocol for months.
Recently, we observed large sheets of debris floating in the culture flasks. Immediately after first noting this debris, we observed that the cells were no longer expanding/growing.
We have replenished our reagents with all new lots/bottles, and re-performed the same experiment, and again, debris was observed.
We have observed this debris in both coated and uncoated flasks, hinting at the possibility that this may be an issue with the flasks. We have also seen this issue before/without the presence of cells, so we have ruled out the cells/vials as the source of the debris.
Images of the debris are attached.
Has anyone ever seen this before in culture? Or have any ideas as to what this may be?
Thanks!
Hello, Laurie!
The pictures seem to show the filaments of the mycelium of the fungus.
They are very difficult to remove. It is necessary to carry out a complete disinfection of reagents, culture dishes and equipment (laminar, incubator).
Hello, Laurie!
The pictures seem to show the filaments of the mycelium of the fungus.
They are very difficult to remove. It is necessary to carry out a complete disinfection of reagents, culture dishes and equipment (laminar, incubator).
Do you use serum-containing medium? Precipitated protein sheets can form in bad batches. That would also affect your rate of cell growth and cell appearance. Common result of too many freeze-thaw cycles. Heat inactivation produces fibrin and lipoprotein precipitates like yours, also.
Does it grow? Yes - possibly fungus [thus it doesn't look like it for me], no- some medium precipitants [scratches of plastic, does it look crystal-shape?].
Try to filter through 0,22um, when it re-appear it is alive.
In both cases, I would consider discarding this particular medium bottle.
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