please help me figure out what's going on with my MDCK cells.
I'm growing MDCK cell in DMEM+10%FCS, and subculture them every other day by trypsinization (0.25%typsin-EDTA, 37C for ~6 min and then remove the trypsin and keep at 37C for another 6 min, hit and tap the flask to detach the cells). It worked well before. but now the cells become very weird. they grow over each other and aggregate, and some are weirdly shaped. they keep dying for several batches. so 2 weeks ago, I switched to MEM+1 mM L-Glutamine+10%FBS. they grew fine for 2 weeks, but were dead again this Monday.
I initially suspected they were contaminated by other virus, because they didn't look like infected by bacteria or mycoplasma. but it turned out to be negative for the other virus kept in our lab (containing GFP, so easy to check for contamination).
And the weirdest thing is that there are other guys in our lab growing the same cell using the same method, and their cells are fine. and all my cells are obtained from them which are fine before getting to my hands. the guy who taught me cell culture watched me doing it and said he can't find anything wrong with my method, but still my cells keep dying while others are fine. It has been torturing me for over 2 months and I can't figure out what's wrong.
I'm attaching photos of the cells. PLEASE PLEASE PLEASE help me! thanks!
Edited by MDCK, 14 June 2011 - 05:52 PM.