Hi all,
I have a somewhat basic question. But as I did not do such experiments before I was hoping someone could provide me with a quick answer whether what I am thinking to do is the correct way.
I have cells for which I need to do a kinetic proliferation assay over 8 weeks to see the effect of drug treatment on their growth. These cells like a particular cell density range and also for some reason grow best in 96 wp wells only. So I need to replate them every week and stick to 96 wp format. They also differentiate on the way in this culture media and most probably will have a differential kinetics at different phases of culture over the duration of experiment. So I want to see not only endpoint cell outputs but also kinetics for different phases,
I was thinking to plate say 100k cells, in a week measure the total cell number, say it is 400k. Doubling coefficient is 4x. I replate them 100k again, next week I get say 350k, the coefficient is 3.5x. So I put on the graph points 100k, 100k*4=400k and 100k*4*3.5=1400k and so on and that will be my growth curve.
I can also transform the cell number axis to a log format. Or I can plot not the cell numbers but doubling coefficient themselves to see how drug treatment affects them every week.
If drug treated cells will be dying more/growing less, I will be still matching the replated cell numbers for treated and control cells every time.
Is this correct?
Hello
Kindly look at link & PDF here
https://www.researchgate.net/post/Long_term_proliferation_assays
Hello!
With this method of analysis, you will have a mixture of dynamics of at least three processes:
1) The dynamics of cell exit from the cell cycle in Ж0 (with or without differentiation)
2) The dynamics of cell cycle deceleration (lengthening J1 mainly)
3) Dynamics of cell survival (necrosis + apoptosis).
If your influence disturbs the mitotic division, then a partial polyploidization of the cells is possible.
You have not written what cells you are researching - attached or cultured in volume.
For attached cells, contact inhibition is also of great importance.
For cell cycle analysis see my publications:
Article Analysis of the Cell Cycle and a Method Employing Synchroniz...
Article Kinetic analysis of cell population growth during culturing in vitro
Hi dear Oleksandr Galkin
I think the following paper can help you;
Best and kind regard.
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