Which is the best way to generate growth curves for cancer cell lines - (Feb/25/2013 )
I am quite new in all these processes and would love some ideas.
I want to generate a growth curves for lung cancer cell lines. They are adhering cells. Do I have to always trypsinize them every 12hrs and count them using a haemocytometer?
Do I seed them on 96 well plates and have different plates for 12hrs. 24hrs. 48hrs. 72hrs and so on and then add MTS reagent to read off the absorbance. I am worried about this though because MTS does not stop the growth so they might increase within the 3hr incubation period. I want to be as accurate as possible.
Thanks you all for your kind response.
you could find many discussions regading growth curve in this forum, just type growth curve in the search box and that will lead you to all those discussions.
I have read them and still unsure. Most of them relate to bacterial cell growth/count
may be this would partially answer your question....
Any method that will give you a measure of the amount of growth should be fine - Assuming a normally cycling population of cells you could measure protein content, DNA content, cell number, cell viability (MTT/MTS assay), ...
Your comment regarding the MTS assay doesn't make sense - over the course of the assay, at every time point you will have to do the same 3h incubation (and same potential 3 hours of growth), so the measures will still be accurate...
Thanks GNANA and bob1.
You can always have duplicates or triplicates for your MTS, I have found it to be a consistent method to assess cell growth. You could also create a standard MTS curve with hemocytometer counts (i.e. you count standard numbers of cells in the beginning, like .5M, 1M, 2M cells) and then you do MTS on those cells. It's useful because then you can skip the hemocytometer counting altogether, which is quite relieving when you have tons of other things to do...
Would be very interested to see a follow up to this discussion if possible! Link removed as irrelevant - bob