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yeast conatamination in primary cells - (Jan/20/2014 )

I am  started a primary hepatocyte cell culture, At the tme of harvest we got around 92% viabile cells and now the cells were maintained in DMEM containing 10% FBS.Now I'm in a doubt about yeast contamination,but no turbidity and color change of the media.But some small particles are floatting and i guess its number is also increasing.But the hepatocytes were finely attached and growing well initially,But now on 8 th day the cell number is too low. I'm attaching the image.Please confirm whether these are yeast contamination or cell debris?I added only pen/srep but no amphptericin.Attached Image

Attached Image

-anju p nair-

It's a little hard to tell as the pictures are too small, but from what I could see, those are most likely debris.  Yeast will show as budding spheres or egg shapes.



Thank you for your reply.How can I get rid of this cell debris during isolation? Whether the cell debris move?

-anju p nair-

Short answer: you can't get rid of it, it is the result of cells dying (hence slow increase), so even if you filter your medium each day, there would still be more appear as the cells die. 


Cell debris can move or it can remain attached.


I agree that this is most likely cellular debris.  But, you can get a ypd plate from a yeast lab and plate a small amount of the media and incubate at 30 to see if colonies grow. You can also take some of the media from these cells and place it in a plate or flask by itself.  See if the particles increase without the cells.  There's no way to completely eliminate cellular debris but you can wash and replace the media daily to minimize.