I wonder if it is possible, in concept, to evaluate the basal phenotype of cell lines, and compare them. For example, If you want to compare the apoptosis potency of cell line A to that of cell line B, how do you normalize the conditions ( growth media, cell number, and cell cycle phase)? how do you select the optimum time point?
Dear Nashed,
You have your answer the question itself, yeas there are number of published articles that you can refer for specific time points, agent used to induce apoptosis, cell cycle arrest etc, also you can standardize the same
For eg. you can do a time kinetics experiment, say you want to study apoptosis in the presence of an agent A, seed cells in a 6 well plate. Either treat them at different time points or you can treat them at the same time and collect them at different time points (say 3, 6,12, 24 and 48 h), I suggest you treat them at different time points and collect them all together (whatever is convenient to you). Do the same stuff with two different cell lines, analyse them with flow cytometry and compare.
Hope this would be hopeful.
Akash Sabarwal Thank you for the answer. However, what i meant in my question is comparing the basal phenotype of two cell lines without treatment or any perturbation. Theoretically, and as the cell lines harbor different genotypes, they must have different phenotypes, at least when the right time point/ condition is captured.
Do the cell lines have a similar morphology, cell size etc.? Are they sub-cell lines where one is derived from another one? Or are they completely independent cell lines?
I would strongly suggest you to culture them under controlled conditions, seed the same cell number in the same amount of medium as if you take them randomly from your normal culture that has maybe a different cell number, different metabolism activity, cells might already behave different. Having the cells in the same cell cycle phase would be perfect. Most times I seed the cells at least for several hours in fresh medium, same cell number, same amount of medium to compare them - but even in different multiple myeloma cell lines, cell size and morphology is very different - along with this RNA, DNA and protein content. Not to talk about the genetic background of different cell lines.
When preparing resistant sub-cell lines I found that you have multiple changes rather than only one particular event (e.g. point mutation as one mechanism of resistance, gene/protein upregulation of the same gene/protein where you have the mutation, but also generally reduced signaling pathway activity, changes in metabolism activity, slower/faster growth).
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