What happen to my NIH3T3 cells? - (Mar/22/2008 )
Hello everyone,
The NIH3T3 were transfected with a gene with unknown function. The transfected cells have strange phenotype. But a small portion of non-transfected cells also have this phenotype. What might be affected in the transfectants?
Most likely it was due to toxicity. Your cell density is on the lower side, therefore the reagent-to-cell ratio is high that can cause toxicity. Probably most will not survive after 24-48 hrs. Try increasing the cell number 2-3 fold, it wont hurt.
Thank you genehunter-1,
But the transgene was delivered by lentivirus. Shown in the pictures is one of the isolated clones.
Sometimes you get sick looking cells if you waited for too long before passing. If so, it will recover after 2-3 passes.
NIH3T3 cells seem to be more sensitive to that than other cell types.
NIH3T3 cells seem to be more sensitive to that than other cell types.
I agree. But after several passages, the cells still appeard like that. And they grow very slowly.
Are they looking normal when you passed them? If both transfected or non-transfected (without any treatment) are looking the same, it may be possible if your cells are not healthy. You can try to thaw a new stock ...
Franz K, thank for your advice.
I think the transgene likely disturb the formation of lamelipodia.