7 replies to this topic

[csegal]

Hi,

I'm culturing adherent LNCaP cells and am concerned that I have contamination - see attached pictures. I contacted HPA, who I bought the cells from, and they suggested that it could be a fault in the plastic (no) or condensation on inside of flask (also no). Does anyone have any better ideas?

Generally, the objects are round, clear looking and larger than the cells. They are floating in the media at the level of the cells (i.e. not air bubbles at the surface).

I sterile filtered all the components that I added to the base media and started fresh reagents. The cells seem to be growing fine and the media doesn't seem to be exhausting quicker than normal.

Suggestions please?

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[Dr Teeth]

csegal, on Oct 9 2009, 11:07 AM, said:

Hi,

I'm culturing adherent LNCaP cells and am concerned that I have contamination - see attached pictures. I contacted HPA, who I bought the cells from, and they suggested that it could be a fault in the plastic (no) or condensation on inside of flask (also no). Does anyone have any better ideas?

Generally, the objects are round, clear looking and larger than the cells. They are floating in the media at the level of the cells (i.e. not air bubbles at the surface).

I sterile filtered all the components that I added to the base media and started fresh reagents. The cells seem to be growing fine and the media doesn't seem to be exhausting quicker than normal.

Suggestions please?

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Are you sure these aren't dead cells?

[CellSpecific.com]

It appears you might be able to wash these floaters away. Try several washes with medium in between aspirations, return to culture and see if the floaters reappear.

[mtrnbh]

Is there a risk of cross contamination with a non-adherent cell line?

[bob1]

dead cells.

[rhombus]

bob1, on Oct 12 2009, 12:50 AM, said:

dead cells.

Agree with bob1, dead cells called "ghost cells"...where most of the intracellular material has been lost and a "ghost membrane" remains.

Rhombus

[eberthella]

Would a live/dead stain help?

[csegal]

Thanks for the suggestions.

I'm pretty convinced it is not cross contamination with another cell line as I've just purchased these from HPA and I'm not culturing any other lines (and I have sole use of the incubator). As for ghost cells, I thought I knew what these looked like and these objects don't look like what I had thought were ghost cells! I'll do a stain and see what happens.

Thanks for the input everyone.