MDCK cell experience - (Aug/04/2008 )

Hi,
Has anyone had any experience with culturing of MDCK cells, in particular high passage numbers? I have MDCKs that have been sub-cultured now to quite a high passage number (>40-50 I think!), however their growth has slowed, but more importantly there are large structures visible down the light microscope that appear to be large vacuoles, bigger than the cells themselves, in the middle of the groups of MDCKs. Thes structures have increased in number and size, either they are gaps between cells or something is being secreted into internal vesicles at a high rate causing these huge vacuolar structures!

I'm going to ditch them and revive new ones, but wondering if anyone has any experience with this, seen this themselves at all/at high passage numbers, or any thoughts as to what this may be/why it has occurred?! Am curious!

Thanks,
mcl

hi, i have the same experience with u too, the MDCK i am currently working on is p26, i see so many big granules under microscope as long as the cells are over ~p20 to p22, and the one who taught me cell culture said this morphology was very normal for MDCK, that was the MDCK can still be used, even though personally i don't prefer to use them.

Hi,
Has anyone had any experience with culturing of MDCK cells, in particular high passage numbers? I have MDCKs that have been sub-cultured now to quite a high passage number (>40-50 I think!), however their growth has slowed, but more importantly there are large structures visible down the light microscope that appear to be large vacuoles, bigger than the cells themselves, in the middle of the groups of MDCKs. Thes structures have increased in number and size, either they are gaps between cells or something is being secreted into internal vesicles at a high rate causing these huge vacuolar structures!

I'm going to ditch them and revive new ones, but wondering if anyone has any experience with this, seen this themselves at all/at high passage numbers, or any thoughts as to what this may be/why it has occurred?! Am curious!

Thanks,
mcl