In my experiment i use tritium labeled thymidine for my cells and incubate for more than 24 hours before assessing the counts per minute as a read out for measuring proliferation of my cells.
Though for other cell lines i get a decent count for healthy cells but for colo-205 I get a very low count even though my cells look healthy and perform well in MTT and annexin assays.
What can be the possible explanations for this?
Thanks.
Hi Moumita,
Does the tritium assay involve a bit more handeling/pipetting? I always disliked doing assays with Colo-205 because they do not attach to the bottom of wells/plates.
Instead of replacing the whole medium to handle the cells, I would remove half carefully and adapt concentrations accordingly (something similar to suspension cultures).
Best,
Noel
Hi Noel,
Post treatment i directly add labeled thymidine to the media already in wells. So i believe this should not be the isuue.
Hi Moumita, Do a trypan blue assay of the same sample and see if you get any variation. Also, the method you are using is dependent on cell division. Viability, Apoptosis are different phenomenon, it might be possible that whatever agent you are using to treat the cells is interfering with DNA replication at a very early stage.