Causes for vacuoles or bubbles in cultured cells - (Jul/23/2008 )

my stock HeLa cells don't look healthy, they are getting some bubble-shaped particles inside. see the attached photos.

why is that? is this due to contamination? high level of serum? or what? they look like fatty particles.

+ to Rhombus,

maybe you're letting them grow too densely before splitting? Cells should be kept growing at log phase, not getting crowded and stalling, then starting up again.

I work with ECC-1 cells (endometrial adenocarcinoma cell line) and they also have these "vacuoles". I know they are not contaminated or over trypsinaized and I use the serum and RPMI from ATCC. What is a too high passage number?

I agree with Rhombus. I think your HeLa cells are 100% confluent (what I see on the picture). It is very possible they are unhappy and that's why they have vacuoles inside. They do not like contaminated.
Usually HeLa cells don't form such vacuoles when they grow happily.

Split your cells when they are 70-80% confluent and so that they will reach the same confluency after 2-4 days.

Yes, actually my cells were too confluent. I think that is the reason why. I usually trypsinize when they are over 90% confluent, so I should avoid that.

they are not contaminated at all. I wash them frequently.

thanks guys.

Thanks for your answer Rhombus. I purchased my cells from ATCC so I went back to their website and checked things out further. I talked to a representative too. These cells are a little atypical. See the picture attached.