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GE1 cell difficult to detach


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#1 sitamanda

sitamanda

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Posted 30 September 2013 - 12:19 AM

Hi!

 

Anyone has any experience on working with GE1 (mouse gingival epithelium) cells?

 

I culture GE1 cells on 9 cm plastic culture plate using 6 ml 1.0 glucose DMEM with 10% FBS and 1% antibiotics. I passage the cells every 2-3 days using 2 ml trypsin-EDTA for 6 minutes, 37 degrees incubation.

My problem is, even after 6 minutes, only half or less cells detached from the plates. I still can see many attached cells. My supervisor said that this is the cell characteristics.

 

I usually just use the detached cells for doing my experiments (checking gene expressions) after seeding them on 24 well plates but I like to keep my original culture plate just in case.

I read that difficult to detach cells are differentiated cells. My supervisor said that GE1 cells are easily differentiated. That's why I tried not to use these strongly attached cells in my experiments.

 

Before, I reseeded my cells on new plate on every passage so I can ensure that every cell that I culture is "good" cells. But, my lab is very "economical" lately. We have to use the same plate until 30 something passages. This frustrate me because the cells cannot be detached completely. I tried longer trypsin-edta incubation, not much difference. I even tried twice trypsin-edta treatment and most of the cells died. (doh)

 

So, I just want to know if anyone has any explanation/experience on this difficult to detach cells. Any suggestion will be appreciated!






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